Heterodera filipjevi continues to be a major threat to wheat production worldwide.Rapid detection and quantification of cyst nematodes are essential for more effective control against this nematode disease.In the pres...Heterodera filipjevi continues to be a major threat to wheat production worldwide.Rapid detection and quantification of cyst nematodes are essential for more effective control against this nematode disease.In the present study,a TaqManminor groove binder(TaqMan-MGB)probe-based fluorescence quantitative real-time PCR(qPCR)was successfully developed and used for quantifying H.filipjevi from DNA extracts of soil.The primers and probe designed from the obtained RAPD-SCAR marker fragments of H.filipjevi showed high specificity to H.filipjevi using DNA from isolatesconfirmed species of 23 Heterodera spp.,1 Globodera spp.and 3 Pratylenchus spp.The qPCR assay is highly sensitive and provides improved H.filipjevi detection sensitivity of as low as 4^(-3) single second-stage juvenile(J2)DNAs,10^(-3) female DNAs,and 0.01μgμL^(-1) genomic DNAs.A standard curve relating to the threshold cycle and log values of nematode numbers was generated and validated from artificially infested soils and was used to quantify H.filipjevi in naturally infested field soils.There was a high correlation between the H.filipjevi numbers estimated from 32 naturally infested field soils by both conventional methods and the numbers quantified using the qPCR assay.qPCR potentially provides a useful platform for the efficient detection and quantification of H.filipjevi directly from field soils and to quantify this species directly from DNA extracts of field soils.展开更多
BACKGROUND Endoscopic ultrasonography-guided fine-needle aspiration(EUS-FNA)and endobronchial ultrasound-guided transbronchial needle aspiration(EBUS-TBNA)are highly sensitive for diagnosing and staging lung cancer.In...BACKGROUND Endoscopic ultrasonography-guided fine-needle aspiration(EUS-FNA)and endobronchial ultrasound-guided transbronchial needle aspiration(EBUS-TBNA)are highly sensitive for diagnosing and staging lung cancer.In recent years,targeted therapy has shown great significance in the treatment of non-small cell lung carcinoma(NSCLC).Using these minimally invasive techniques to obtain specimens for molecular testing will provide patients with a more convenient diagnostic approach.AIM To evaluate the feasibility and accuracy of tissue samples obtained using EUSFNA and EBUS-TBNA for molecular diagnosis of NSCLC.METHODS A total of 83 patients with NSCLC underwent molecular testing using tissues obtained from EUS-FNA or EBUS-TBNA at the Tianjin Medical University Cancer Hospital from January 2017 to June 2019.All enrolled patients underwent chest computed tomography or positron emission tomography/computed tomography prior to puncture.We detected abnormal expression of EGFR,KRAS,MET,HER2,ROS1 and anaplastic lymphoma kinase protein.Two patients failed to complete molecular testing due to insufficient tumor tissue.The clinical features,puncture records,molecular testing results and targeted treatment in the remaining 81 patients were summarized.RESULTS In a total of 99 tissue samples obtained from 83 patients,molecular testing was successfully completed in 93 samples with a sample adequacy ratio of 93.9%(93/99).Biopsy samples from two patients failed to provide test results due to insufficient tumor tissue.In the remaining 81 patients,62 cases(76.5%)were found to have adenocarcinoma,11 cases(13.6%)had squamous cell carcinoma,3 cases(3.7%)had adenosquamous carcinoma and 5 cases(6.2%)had NSCLC-not otherwise specified.The results of molecular testing showed EGFR mutations in 21 cases(25.9%),KRAS mutations in 9 cases(11.1%),ROS-1 rearrangement in 1 case(1.2%)and anaplastic lymphoma kinase-positive in 5 cases(6.2%).Twentyfour patients with positive results received targeted therapy.The total effectiveness rate of targeted therapy was 66.7%(16/24),and the disease control rate was 83.3%(20/24).CONCLUSION Tissue samples obtained by EUS-FNA or EBUS-TBNA are feasible for the molecular diagnosis of NSCLC and can provide reliable evidence for clinical diagnosis and treatment.展开更多
Hearing loss is one of the most common birth defects,with inherited genetic defects play an important role,contributing to about 60%of deafness occurring in infants.However,hearing impairment is genetically heterogene...Hearing loss is one of the most common birth defects,with inherited genetic defects play an important role,contributing to about 60%of deafness occurring in infants.However,hearing impairment is genetically heterogeneous,with both common and rare forms occurring due to mutations in estimated 500 genes.Due to the large number and presumably low mutation frequencies of those genes,it would be highly expensive and time-consuming to address this issue by conventional gene-by-gene Sanger sequencing.Next-generation sequencing is a revolutionary technology that allows the simultaneous screening of mutations in a large number of genes.It is cost effective compared to classical strategies of linkage analysis and direct sequencing when the number or size of genes is large,and thus has become a highly efficient strategy for identifying novel causative genes and mutations involved in heritable disease.In this review,we describe major NGS methodologies currently used for genetic disorders and highlight applications of these technologies in studies of molecular diagnosis and the discovery of genes implicated in non-syndromic hearing loss.展开更多
Background The complexity of the Chagas disease and its phases is impossible to have a unique test for both phases and a lot of different epidemiological scenarios.Currently,serology is the reference standard techniqu...Background The complexity of the Chagas disease and its phases is impossible to have a unique test for both phases and a lot of different epidemiological scenarios.Currently,serology is the reference standard technique;occasionally,results are inconclusive,and a different diagnostic technique is needed.Some guidelines recommend molecular testing.A systematic review and meta-analysis of available molecular tools/techniques for the diagnosis of Chagas disease was performed to measure their heterogeneity and efficacy in detecting Trypanosoma cruzi infection in blood samples.Methods A systematic review was conducted up to July 27,2022,including studies published in international databases.Inclusion and exclusion criteria were defined to select eligible studies.Data were extracted and presented according to PRISMA 2020 guidelines.Study quality was assessed using Quality Assessment of Diagnostic Accuracy Studies-2(QUADAS-2).A random-effects model was used to calculate pooled sensitivity,specificity,and diagnostic odds ratio(DOR).Forest plots and a summary of the receiving operating characteristics(SROC)curves displayed the outcomes.Heterogeneity was determined by I^(2)and Tau^(2)statistics and P values.Funnel plots and Deek's test were used to assess publication bias.A quantitative meta-analysis of the different outcomes in the two different clinical phases was performed.Results We identified 858 records and selected 32 papers.Studies pertained to endemic countries and nonendemic areas with adult and paediatric populations.The sample sizes ranged from 17 to 708 patients.There were no concerns regarding the risk of bias and applicability of all included studies.A positive and nonsignificant correlation coefficient(S=0.020;P=0.992)was obtained in the set of studies that evaluated diagnostic tests in the acute phase population(ACD).A positive and significant correlation coefficient(S=0.597;P<0.000)was obtained in the case of studies performed in the chronic phase population(CCD).This resulted in high heterogeneity between studies,with the master mix origin and guanidine addition representing significant sources.Interpretation/Conclusions and relevance The results described in this meta-analysis(qualitative and quantitative analyses)do not allow the selection of the optimal protocol of molecular method for the study of Trypanosoma cruzi infection in any of its phases,among other reasons due to the complexity of this infection.Continuous analysis and optimization of the different molecular techniques is crucial to implement this efficient diagnosis in endemic areas.展开更多
Hepatocellular carcinoma(HCC)is often associated with pre-existing chronic liver pathologies of different origin infections of hepatitis B virus(HBV)and hepatitis C virus.Clinically,the diagnosis and therapy for HCC a...Hepatocellular carcinoma(HCC)is often associated with pre-existing chronic liver pathologies of different origin infections of hepatitis B virus(HBV)and hepatitis C virus.Clinically,the diagnosis and therapy for HCC are very important for the prognosis of patients.However,current methods for HCC diagnosis and therapy have no an optimal accuracy due to the tumor heterogeneity and the frequent late diagnosis.This review summarizes the new advances in molecular diagnosis and therapy of HCC,based on the recent novel biomarkers and new therapeutic strategies for HCC,including alpha-fetoprotein-L3,glypican-3,heat shock protein 90,dickkopf WNT signaling pathway inhibitor 1,paraoxonase 1,highly up-regulated in liver cancer.Moreover,epigenetic regulation,signal pathway,cellular and molecular targets for the immunotherapy,tumor microenviroment and genome sequencing analysis may serve as the molecular expression signatures in clinical practice.For promising new treatment strategy of HCC,targeting molecular therapy based on the restoration of tumor suppressor genes lost and inhibition of oncogenic genes is attractive.The new clinical trials for other molecular-targeted agents,including pembrolizumab,nivolumab,tivantinib,lenvatinib,cabozantinib,and ramucirumab,are ongoing in clinic.Interestingly,anti-HBV drugs display an amazing therapy for HBV-related HCC.In future,the global determination of more biomarkers may provide new insights into the diagnosis of HCC.More importantly,the diagnostic markers should be used to trace patient's follow-up disease progression,guiding doctors to judge and prescribe drugs for status of an illness,prognosis and other processes.展开更多
Supersonic Molecular Beam Injection (SMBI) is a new fuelling method for Tokamaks and has recently been improved to enhance the flux of the beam and to make a survey of the cluster effect within the beam. There are a s...Supersonic Molecular Beam Injection (SMBI) is a new fuelling method for Tokamaks and has recently been improved to enhance the flux of the beam and to make a survey of the cluster effect within the beam. There are a series of new phenomena, which implicate the interaction of the beam (including clusters) with the toroidal plasma of HL-1M Tokamak. The Ha signals from the edge show a regular variation around the torus. Around the injection port, the edge Hα signals are positive rectangular wave, which is consistent with that of the injection beam pulses. The edge electron temperature, measured with movable Langmuir probes, decreases by an order of magnitude and the density increases by an order of magnitude. Hα emission at the beam injection port, measured with CCD camera at an angle of 13.4 degrees to the SMBI line, shows many separate peaks within the contour plot. These peaks may show the strong emission produced by the interaction of the hydrogen clusters with the plasma. Hydrogen clusters may be produced in the beam according to the empirical scaling (Hagena) law of clustering onset, * = .here d is the nozzle diameter in μm, Po the stagnation pressure in mbar, To the source temperature in K, and k is a constant related to the gas species. If * > 100, clusters will be formed. In present experiment * is about 127.展开更多
Objective:To determine an algorithm for molecular diagnosis of visceral leishmaniasis(VL)by kinetoplast DNA(kDNA)(RV1/RV2)and internal transcriber spacer(ITS1)(LITSR/L5.8 S)polymerase chain reaction(PCR),complemented ...Objective:To determine an algorithm for molecular diagnosis of visceral leishmaniasis(VL)by kinetoplast DNA(kDNA)(RV1/RV2)and internal transcriber spacer(ITS1)(LITSR/L5.8 S)polymerase chain reaction(PCR),complemented by ITS 1 PCR restriction fragment length polymorphism(RFLP),using peripheral blood or bone marrow aspirate from patients with suspected VL.Methods:Biological samples were submitted to the gold standard for the diagnosis of VL and molecular diagnosis represented by ITS 1 PCR,kDNA PCR,and ITS 1 PCR RFLP.The samples were obtained from seven groups:groupⅠ,82 samples from patients with confirmed VL;groupⅡ,16 samples from patients under treatment for VL;groupⅢ,14 samples from dogs with canine visceral leishmaniasis(CVL);groupⅣ,a pool of six experimentally infected sandflies(Lutzomya longipalpis);group V,18 samples from patients with confirmed tegumentary leishmaniasis(TL)and groupsⅥandⅦwere from control groups without VL.Results:The following gold standard and molecular examination results were obtained for each of the seven groups:groupⅠ:parasitologic and immunochromatographic tests showed a sensitivity of 76.3%(61 of 80)and 68.8%(55 of 80),respectively,and a sensitivity of 97.6%(80 of 82)and 92.7%(76 of 82)by ITS1 and kDNA PCR,respectively.After ITS1 PCR RFLP(HaeⅢ)analysis of the 80 positive samples,52.5%(42 of 80)generated three fragments of 180,70,and 50 bp,corresponding to the pattern of Leishmania infantum infantum;groupⅡ:negative for the parasitologic methods and positive for IrK39(100%,16 of 16),presented 12.5%(2 of 16)of positivity by ITS 1 PCR and 25.0%(4 of 16)by kDNA PCR;groupⅢ:positive in the parasitologic and serologic tests(100%,14 of 14),presented 85.7%(12 of 14)of positivity by ITS1 PCR and kDNA PCR.ITS1 PCR RFLP showed that 83.3%(10 of 12)of the canine samples contained parasites with profiles similar to L.infantum;groupⅣpresented amplifications by ITS1 PCR and kDNA PCR.ITS1 PCR products were analyzed by RFLP,generating a profile similar to that of L.infantum;groupⅤ:positive in the parasitologic examination(100%,18 of 18),presented 72.2%(13 of 18)of the samples by ITS1 PCR positive.A total of 69.2%(9 of 13)showed profiles corresponding to a Viannia complex by ITS1 PCR RFLP;and groupⅥand groupⅥwere negative by ITS 1 and kDNA molecular tests.Comparing the molecular results with the parasitologic and serologic diagnosis from groupⅠ,almost perfect agreement was found(κboth>0.80,P<0.001).ITS1 and RV1/RV2 PCR detected 90.2%(74 of 82)of the samples.Two samples positive by RV1/RV2 were negative by LITSR/L5.8 S,and six samples positive by LITSR/L5.8 S were negative by RV1/RV2.Therefore,these two systems complemented each other;they diagnosed 100%of the samples as belonging to the Leishmania genus.Conclusions:We suggest an algorithm for the molecular diagnosis of VL,which must consider previous parasitologic and serologic(immunochromatographic)diagnoses,and should combine kDNA and ITS1 to determine the Leishmania subgenus using RFLP as a complement method to define the L.infantum species.展开更多
The existence of an inflammatory process in the heart muscle,related to a progressive worsening of myocardial function,different etiopathogenetic mechanisms concur and often overlap,thus making the diagnosis and the t...The existence of an inflammatory process in the heart muscle,related to a progressive worsening of myocardial function,different etiopathogenetic mechanisms concur and often overlap,thus making the diagnosis and the therapeutic approach complex.As the COVID-19 pandemic progresses,the effects of the disease on the organ systems and in particular on the cardiovascular system are becoming more and more profound.Cardiac involvement is a well-known event with a high percentage of findings in the heart’s magnetic field,even in asymptomatic areas.There are numerous uncertainties regarding their evolution,in the long and short term,due not only to a difficult to determine the varied clinical expression and the rarely performed intramyocardial biopsy which additionally presents diagnostic problems but also in part to different clinical prognosis.Today,the new SARS-CoV-2 virus that uses the angiotensin converting enzyme 2(ACE2)which is present at high levels in myocardial cells as its entrance it can create even severe heart injury.The pathophysiology in all of these cases can involve multiple immune and non-immune mechanisms within organs and vessels and can be occur in the clinical phases.Possible mechanisms of direct and indirect myocardial infarction in patients with COVID-19 include additional lesion and oxygen-rich and generalized inflammation response with myocardial immune hyperactivity(myocarditis).Therefore,these can occur through the excessive release of cytokines,the presence of thrombocytopenia,endocrine damage,heart failure,arrhythmias and more.Patients can show average signs of myocardial damage,and some develop spontaneous cardiac complications,such as heart failure,arrhythmias and,rarely,rare cardiogenic disorders.Pathophysiology in all of these may involve multiple mechanisms within the cytokine cephalic membrane,endocrine damage and thrombogenicity.The diagnosis of this myocardial injuri is mainly based on the myocardial enzyme troponin.This viewpoint paper explains today’s knowledge on viral myocarditis,in particular that from SARS-CoV-2 infection,if there is a connection with other possible biomolecular pathogenetic factors that can influence its natural course.In fact,it is for this reason that the pathogenetic mechanisms are analyzed and described.At the same time,its possible interaction with other parameters that are documented risk factors for cardiovascular disease was examined.Although these biomolecular findings were mainly related to necrotic parts of the myocardium,it is important to recognize that myocardial damage early for a better approach and prognosis.展开更多
<b><span style="font-family:Verdana;">Background:</span></b><span style="font-family:Verdana;"> Extrapulmonary tuberculosis (EPTB) remains difficult to diagnose becaus...<b><span style="font-family:Verdana;">Background:</span></b><span style="font-family:Verdana;"> Extrapulmonary tuberculosis (EPTB) remains difficult to diagnose because the clinical specimens to be examined are often paucibacillary</span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;"> and obtained with difficulty from inaccessible sites. </span><span style="font-family:Verdana;">An updated Xpert<sup></sup></span></span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"><sup><span style="white-space:nowrap;">®</span></sup></span></span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> MTB/RIF Ultra (Ultra) test has been designed and licensed to improve sensitivity in the detection of Mycobacterium tuberculosis complex.</span></span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;"> The aim of the present study is to evaluate the performance of Ultra assay for the clinical diagnosis of EPTB in </span><span style="font-family:Verdana;">a low tuberculosis prevalence country. </span><span style="font-family:Verdana;"><b></b></span><b><b><span style="font-family:Verdana;">Methods:</span></b><span style="font-family:Verdana;"></span></b></span><span style="font-family:Verdana;"> A retrospective analysis was performed at “A.</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">O dei Colli” of Naples on consecutive extrapulmonary specimens for EPTB across a three-year period. All different types of extrapulmonary specimens were tested for EPTB by smear microscopy, culture and Ultra assay in accordance with relevant guidelines. </span><span style="font-family:Verdana;"><b></b></span><b><b><span style="font-family:Verdana;">Results:</span></b><span style="font-family:Verdana;"></span></b></span><b> </b><span style="font-family:Verdana;">A total of 606 EPTB samples, 561 culture negative EPTB and 45 culture positive EPTB were included. Using culture as reference standard, the overall sensitivities and specificities of Ultra assay were 95.6% (95% CI 84.8</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">-</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">99.5) and 97.5% (95% CI 95.8</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">-</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">98.6) respectively. Sensitivity and specificity of Ultra for individual category of specimens w</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">ere</span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;"> also</span><span style="color:red;"> </span><span style="font-family:Verdana;">performed. </span><span style="font-family:Verdana;"><b></b></span><b><b><span style="font-family:Verdana;">Conclusion:</span></b><span style="font-family:Verdana;"></span></b></span><span style="font-family:Verdana;"> In a </span><span style="font-family:Verdana;">low-tuberculosis prevalence setting, Ultra assay confirms to have a good performance in the diagnosis of EPTB for all different extrapulmonary samples.</span></span></span>展开更多
Background: African animal trypanosomiasis (AAT) is caused mainly by Trypanosoma congolense, T. vivax, and T. brucei brucei and is the major constraint for livestock productivity in Sub-Saharan African countries. Info...Background: African animal trypanosomiasis (AAT) is caused mainly by Trypanosoma congolense, T. vivax, and T. brucei brucei and is the major constraint for livestock productivity in Sub-Saharan African countries. Information about animal trypanosomiasis status in Ivory Coast is missing, especially regarding molecular epidemiology. Therefore, this study intended to apply molecular tools to identify and characterize trypanosomes in Ivory Coast for sustainable control. Methods: 363 cattle blood samples were collected from Ferkessedougou Region in northern Ivory Coast in 2012. Buffy coat technique (BCT) and species-specific PCR assays were used to detect trypanosome species. Results: Out of 363 cattle examined with BCT, 33 were found positive with all trypanosomes species accounting for an average of 9.09% prevalence whereas polymerase chain reaction (PCR) using species-specific primers showed that 81 out of 363 cattle were infected with trypanosomes with an overall prevalence of 22.31%. Trypanosoma congolense savanah type, T. Vivax and T. brucei sl. accounted for 28.39%, 49.38% and 23.45% of the infection rate respectively. No infection with T. congo forest?type was detected. T. vivax infection was the most prevalence in the area investigated compared to the two other trypanosome species. Mixed infections with different trypanosomes species were detected accounting for 7.32% of prevalence. Regarding sexrelated prevalence, male cattles were slightly more infected than female but the difference was not significant. Conclusion: Our results showed that there was a high prevalence of AAT in livestock in Ferkessedougou Area. There is therefore a need to strengthen control policies and institute measures that help prevent the spread of the parasites for sustainable control of animal trypanosome in this area.展开更多
Haemochromatosis is a genetic disease caused by hepcidin deficiency,responsible for an increase in intestinal iron absorption.Haemochromatosis is associated with homozygosity for the HFE p.Cys282Tyr mutation.However,r...Haemochromatosis is a genetic disease caused by hepcidin deficiency,responsible for an increase in intestinal iron absorption.Haemochromatosis is associated with homozygosity for the HFE p.Cys282Tyr mutation.However,rare cases of haemochromatosis(non-HFE haemochromatosis)can also be caused by path-ogenic variants in other genes(such as HJV,HAMP,TFR2 and SLC40A1).A working group of the International Society for the Study of Iron in Biology and Medicine(BIOIRON Society)has concluded that the classification based in different molecular subtypes is difficult to be adopted in clinical practice and has proposed a new classification approaching clinical questions and molecular complexity.The aim of the present review is to provide an update on classification,pathophysiology and therapeutic recommendations.展开更多
Fluorescence imaging is a noninvasive and dynamic real-time imaging technique;however,it exhibits poor spatial resolution in centimeter-deep tissues because biological tissues are highly scattering media for optical r...Fluorescence imaging is a noninvasive and dynamic real-time imaging technique;however,it exhibits poor spatial resolution in centimeter-deep tissues because biological tissues are highly scattering media for optical radiation.The recently developed ultrasound-controlled fluorescence(UCF)imaging is a novel imaging technique that can overcome this bottleneck.Previous studies suggest that the effective contrast agent and sensitive imaging system are the two pivotal factors for generating high-resolution UCF images ex vivo and/or in vivo.Here,this review highlights the recent advances(2015e2020)in the design and synthesis of contrast agents and the improvement of imaging systems to realize high-resolution UCF imaging of deep tissues.The imaging performances of various UCF systems,including the signal-to-noise ratio,imaging resolution,and imaging depth,are specifically discussed.In addition,the challenges and prospects are highlighted.With continuously increasing research interest in this field and emerging multidisciplinary applications,UCF imaging with higher spatial resolution and larger imaging depth may be developed shortly,which is expected to have a far-reaching impact on disease surveillance and/or therapy.展开更多
Objective:To develop a new technique for diagnosis of Plasmodium knowlesi and at the same time to be able to discriminate among the diverse species of Plasmodium causing human malaria.Methods:In this study the nested ...Objective:To develop a new technique for diagnosis of Plasmodium knowlesi and at the same time to be able to discriminate among the diverse species of Plasmodium causing human malaria.Methods:In this study the nested multiplex malaria PCR was redesigned,targeting the 18S rR NA gene,to identify the fifth human Plasmodium species,Plasmodium knowlesi,together with the other human Plasmodium(Plasmodium falciparum,Plasmodium vivax,Plasmodium ovale and Plasmodium malariae)by amplified fragment size using only two amplification processes and including an internal reaction control to avoid false negatives.Results:The technique was validated with 91 clinical samples obtained from patients with malaria compatible symptoms.The technique showed high sensitivity(100%)and specificity(96%)when it was compared to the reference method employed for malaria diagnosis in the Instituto de Salud Carlos栿and a published real-time PCR malaria assay.Conclusions:The technique designed is an economical,sensitive and specific alternative to current diagnosis methods.Furthermore,the method might be tested in knowlesi-malaria endemic areas with a higher number of samples to confirm the quality of the method.展开更多
Objective: The goal of this study was to determine the prevalence of C. trachomatis in women diagnosed with infertility attending the Outpatient Clinic of Infertility from Botucatu Medical School, UNESP, Brazil. Patie...Objective: The goal of this study was to determine the prevalence of C. trachomatis in women diagnosed with infertility attending the Outpatient Clinic of Infertility from Botucatu Medical School, UNESP, Brazil. Patients and Methods: This molecular study enrolled a total of 112 women. Among these patients, 62 presented primary infertility while 50 presented secondary infertility. The criteria for eligibility included women who were: reproductive-aged;no prior report of seroconversion for HIV;no antibiotic or vaginal cream used in the preceding 30 days;and abstinence from sexual intercourse for 72 hours before the visit. The women were submitted to a gynecological examination and cervical samples were collected with an endocervical cytobrush for molecular analysis of C. trachomatis. Results: The prevalence of chlamydial infection was 8% with similar prevalence between primary (8.1%) and secondary (8.0%) infertility. Conclusion: Considering the asymptomatic nature of chlamydial infection and its association with tubal factor infertility, there is a pressing need to incorporate the screening of C. trachomatis infection as part of the routine investigation for infertility. The early diagnostic by screening can minimize complications and reduce Public Health costs with Assisted Reproductive Technology.展开更多
Hepatocellular carcinoma(HCC)is the most common type of primary hepatocellular carcinoma(PHC).Early diagnosis of HCC remains the key to improve the prognosis.In recent years,with the promotion of the concept of precis...Hepatocellular carcinoma(HCC)is the most common type of primary hepatocellular carcinoma(PHC).Early diagnosis of HCC remains the key to improve the prognosis.In recent years,with the promotion of the concept of precision medicine and more in-depth analysis of the biological mechanism underlying HCC,new diagnostic methods,including emerging serum markers,liquid biopsies,molecular diagnosis,and advances in imaging(novel contrast agents and radiomics),have emerged one after another.Herein,we reviewed and analyzed scientific advances in the early diagnosis of HCC and discussed their application and shortcomings.This review aimed to provide a reference for scientific research and clinical practice of HCC.展开更多
Diagnosis of tuberculosis can be difficult as advances in molecular diagnosis approaches(especially nanoparticles combined with high-throughput mass spectrometry for detecting mycobacteria peptide)and personalized med...Diagnosis of tuberculosis can be difficult as advances in molecular diagnosis approaches(especially nanoparticles combined with high-throughput mass spectrometry for detecting mycobacteria peptide)and personalized medicine result in many changes to the diagnostic framework.This review will address issues concerning novel technologies from bench to bed and new strategies for personalized tuberculosis diagnosis.展开更多
The worldwide epidemic of coronavirus disease 2019(COVID-19)is ongoing.Rapid and accurate detection of the causative virus SARSCoV-2 is vital for the treatment and control of COVID-19.In this study,the comparative sen...The worldwide epidemic of coronavirus disease 2019(COVID-19)is ongoing.Rapid and accurate detection of the causative virus SARSCoV-2 is vital for the treatment and control of COVID-19.In this study,the comparative sensitivity of different respiratory specimen types were retrospectively analyzed using 3,552 clinical samples from 410 COVID-19 patients confirmed by Guangdong CDC(Center for Disease Control and Prevention).Except for bronchoalveolar lavage fluid(BALF),the sputum possessed the highest positive rate(73.4%–87.5%),followed by nasal swabs(53.1%–85.3%)for both severe and mild cases during the first 14 days after illness onset(d.a.o.).Viral RNA could be detected in all BALF samples collected from the severe group within 14 d.a.o.and lasted up to 46 d.a.o.Moreover,although viral RNA was negative in the upper respiratory samples,it was also positive in BALF samples in most cases from the severe group during treatment.Notably,no viral RNA was detected in BALF samples from the mild group.Despite typical ground-glass opacity observed via computed tomographic scans,no viral RNA was detected in the first three or all upper respiratory tract specimens from some COVID-19 patients.In conclusion,sputum is most sensitive for routine laboratory diagnosis of COVID-19,followed by nasal swabs.Detection of viral RNA in BALF improves diagnostic accuracy in severe COVID-19 patients.展开更多
Many seizures in neonates are due to early-onset epilepsy,which is often difficult to diagnose,especially to explore the causes.Recently,the development of next-generation sequencing(NGS)has led to the discovery of a ...Many seizures in neonates are due to early-onset epilepsy,which is often difficult to diagnose,especially to explore the causes.Recently,the development of next-generation sequencing(NGS)has led to the discovery of a large number of genes involved in epilepsy.This may improve prompt detection of early-onset epilepsy in neonates.This study aimed at analyzing the genotype-phenotype correlations in neonates with seizures in a bid to improve the understanding of genetic diagnosis of early-onset epilepsy.Clinical features and prognosis of 15 children who underwent genetic testing having had unexplained seizures from February 2016 to May 2018 in Children’s Hospital of Chongqing Medical University were analyzed retrospectively.The salient findings were:poor response to stimulus and abnormal electroencephalogram(EEG)in the initial period were observed in the group with concomitant genetic abnormalities.Despite the recent progress in genetic technology,molecular diagnosis for neonatal-onset epilepsy can be challenging due to genetic and phenotypic heterogeneities.However,some genotypes are associated with specific clinical manifestations and EEG patterns.Therefore,in-depth understanding of genotype-phenotype correlations would be useful to clinicians managing neonates with early-onset seizures.展开更多
Congenital heart disease(CHD)is the most common birth defect worldwide.In recent years,the widespread application of innovative molecular diagnostic technologies in clinical scenarios has obviously increased the molec...Congenital heart disease(CHD)is the most common birth defect worldwide.In recent years,the widespread application of innovative molecular diagnostic technologies in clinical scenarios has obviously increased the molecular diagnostic yields of CHD,providing evidence-based guidance for medical decision-making.These molecular diagnostic technologies include chromosome microarray analysis,targeted sequencing,exome sequencing,and genome sequencing.Furthermore,high-throughput sequencing technology has performed excellently in the clinical molecular diagnosis of CHD.This review provides an overview of the current technology and applications in the molecular diagnosis of CHD.The unmet issues and future directions in adapting novel genomic testing technologies to the molecular diagnosis of CHD in clinical settings are also addressed.展开更多
A small fraction of patients diagnosed with obesity or diabetes mellitus has an underlying monogenic cause.Here,we constructed a targeted gene panel consisting of 83 genes reported to be causative for monogenic obesit...A small fraction of patients diagnosed with obesity or diabetes mellitus has an underlying monogenic cause.Here,we constructed a targeted gene panel consisting of 83 genes reported to be causative for monogenic obesity or diabetes.We performed this panel in 481 patients to detect causative variants and compared these results with whole-exome sequencing(WES)data available for 146 of these patients.The coverage of targeted gene panel sequencing was significantly higher than that of WES.The diagnostic yield in patients sequenced by the panel was 32.9%with subsequent WES leading to three additional diagnoses with two novel genes.In total,178 variants in 83 genes were detected in 146 patients by targeted sequencing.Three of the 178 variants were missed by WES,although the WES-only approach had a similar diagnostic yield.For the 335 samples only receiving targeted sequencing,the diagnostic yield was 32.2%.In conclusion,taking into account the lower costs,shorter turnaround time,and higher quality of data,targeted sequencing is a more effective screening method for monogenic obesity and diabetes compared to WES.Therefore,this approach could be routinely established and used as a first-tier test in clinical practice for specific patients.展开更多
基金financially supported by the National Natural Science Foundation of China(31972247)the Science and Technology Innovation Project of the Chinese Academy of Agricultural Sciences(ASTIP-2016-IPP-04)the Special Fund for Agro-scientific Research in the Public Interest,China(201503114)。
文摘Heterodera filipjevi continues to be a major threat to wheat production worldwide.Rapid detection and quantification of cyst nematodes are essential for more effective control against this nematode disease.In the present study,a TaqManminor groove binder(TaqMan-MGB)probe-based fluorescence quantitative real-time PCR(qPCR)was successfully developed and used for quantifying H.filipjevi from DNA extracts of soil.The primers and probe designed from the obtained RAPD-SCAR marker fragments of H.filipjevi showed high specificity to H.filipjevi using DNA from isolatesconfirmed species of 23 Heterodera spp.,1 Globodera spp.and 3 Pratylenchus spp.The qPCR assay is highly sensitive and provides improved H.filipjevi detection sensitivity of as low as 4^(-3) single second-stage juvenile(J2)DNAs,10^(-3) female DNAs,and 0.01μgμL^(-1) genomic DNAs.A standard curve relating to the threshold cycle and log values of nematode numbers was generated and validated from artificially infested soils and was used to quantify H.filipjevi in naturally infested field soils.There was a high correlation between the H.filipjevi numbers estimated from 32 naturally infested field soils by both conventional methods and the numbers quantified using the qPCR assay.qPCR potentially provides a useful platform for the efficient detection and quantification of H.filipjevi directly from field soils and to quantify this species directly from DNA extracts of field soils.
基金Supported by National Natural Science Foundation of China,No.81903055Tumor Translational Medicine Seed Fund of Tianjin Medical University Cancer Institute and Hospital,No.1709.
文摘BACKGROUND Endoscopic ultrasonography-guided fine-needle aspiration(EUS-FNA)and endobronchial ultrasound-guided transbronchial needle aspiration(EBUS-TBNA)are highly sensitive for diagnosing and staging lung cancer.In recent years,targeted therapy has shown great significance in the treatment of non-small cell lung carcinoma(NSCLC).Using these minimally invasive techniques to obtain specimens for molecular testing will provide patients with a more convenient diagnostic approach.AIM To evaluate the feasibility and accuracy of tissue samples obtained using EUSFNA and EBUS-TBNA for molecular diagnosis of NSCLC.METHODS A total of 83 patients with NSCLC underwent molecular testing using tissues obtained from EUS-FNA or EBUS-TBNA at the Tianjin Medical University Cancer Hospital from January 2017 to June 2019.All enrolled patients underwent chest computed tomography or positron emission tomography/computed tomography prior to puncture.We detected abnormal expression of EGFR,KRAS,MET,HER2,ROS1 and anaplastic lymphoma kinase protein.Two patients failed to complete molecular testing due to insufficient tumor tissue.The clinical features,puncture records,molecular testing results and targeted treatment in the remaining 81 patients were summarized.RESULTS In a total of 99 tissue samples obtained from 83 patients,molecular testing was successfully completed in 93 samples with a sample adequacy ratio of 93.9%(93/99).Biopsy samples from two patients failed to provide test results due to insufficient tumor tissue.In the remaining 81 patients,62 cases(76.5%)were found to have adenocarcinoma,11 cases(13.6%)had squamous cell carcinoma,3 cases(3.7%)had adenosquamous carcinoma and 5 cases(6.2%)had NSCLC-not otherwise specified.The results of molecular testing showed EGFR mutations in 21 cases(25.9%),KRAS mutations in 9 cases(11.1%),ROS-1 rearrangement in 1 case(1.2%)and anaplastic lymphoma kinase-positive in 5 cases(6.2%).Twentyfour patients with positive results received targeted therapy.The total effectiveness rate of targeted therapy was 66.7%(16/24),and the disease control rate was 83.3%(20/24).CONCLUSION Tissue samples obtained by EUS-FNA or EBUS-TBNA are feasible for the molecular diagnosis of NSCLC and can provide reliable evidence for clinical diagnosis and treatment.
基金supported by grants from the Project of the National Natural Science Foundation of China(Grant Nos.30801285,81230020,81200751,81070792,81000415, 81360159)grants from China Postdoctoral Science Foundation(No.2012M,2013T52187860947)a grant from Minister of Science and Technology of China(2012BAI09B02)
文摘Hearing loss is one of the most common birth defects,with inherited genetic defects play an important role,contributing to about 60%of deafness occurring in infants.However,hearing impairment is genetically heterogeneous,with both common and rare forms occurring due to mutations in estimated 500 genes.Due to the large number and presumably low mutation frequencies of those genes,it would be highly expensive and time-consuming to address this issue by conventional gene-by-gene Sanger sequencing.Next-generation sequencing is a revolutionary technology that allows the simultaneous screening of mutations in a large number of genes.It is cost effective compared to classical strategies of linkage analysis and direct sequencing when the number or size of genes is large,and thus has become a highly efficient strategy for identifying novel causative genes and mutations involved in heritable disease.In this review,we describe major NGS methodologies currently used for genetic disorders and highlight applications of these technologies in studies of molecular diagnosis and the discovery of genes implicated in non-syndromic hearing loss.
文摘Background The complexity of the Chagas disease and its phases is impossible to have a unique test for both phases and a lot of different epidemiological scenarios.Currently,serology is the reference standard technique;occasionally,results are inconclusive,and a different diagnostic technique is needed.Some guidelines recommend molecular testing.A systematic review and meta-analysis of available molecular tools/techniques for the diagnosis of Chagas disease was performed to measure their heterogeneity and efficacy in detecting Trypanosoma cruzi infection in blood samples.Methods A systematic review was conducted up to July 27,2022,including studies published in international databases.Inclusion and exclusion criteria were defined to select eligible studies.Data were extracted and presented according to PRISMA 2020 guidelines.Study quality was assessed using Quality Assessment of Diagnostic Accuracy Studies-2(QUADAS-2).A random-effects model was used to calculate pooled sensitivity,specificity,and diagnostic odds ratio(DOR).Forest plots and a summary of the receiving operating characteristics(SROC)curves displayed the outcomes.Heterogeneity was determined by I^(2)and Tau^(2)statistics and P values.Funnel plots and Deek's test were used to assess publication bias.A quantitative meta-analysis of the different outcomes in the two different clinical phases was performed.Results We identified 858 records and selected 32 papers.Studies pertained to endemic countries and nonendemic areas with adult and paediatric populations.The sample sizes ranged from 17 to 708 patients.There were no concerns regarding the risk of bias and applicability of all included studies.A positive and nonsignificant correlation coefficient(S=0.020;P=0.992)was obtained in the set of studies that evaluated diagnostic tests in the acute phase population(ACD).A positive and significant correlation coefficient(S=0.597;P<0.000)was obtained in the case of studies performed in the chronic phase population(CCD).This resulted in high heterogeneity between studies,with the master mix origin and guanidine addition representing significant sources.Interpretation/Conclusions and relevance The results described in this meta-analysis(qualitative and quantitative analyses)do not allow the selection of the optimal protocol of molecular method for the study of Trypanosoma cruzi infection in any of its phases,among other reasons due to the complexity of this infection.Continuous analysis and optimization of the different molecular techniques is crucial to implement this efficient diagnosis in endemic areas.
文摘Hepatocellular carcinoma(HCC)is often associated with pre-existing chronic liver pathologies of different origin infections of hepatitis B virus(HBV)and hepatitis C virus.Clinically,the diagnosis and therapy for HCC are very important for the prognosis of patients.However,current methods for HCC diagnosis and therapy have no an optimal accuracy due to the tumor heterogeneity and the frequent late diagnosis.This review summarizes the new advances in molecular diagnosis and therapy of HCC,based on the recent novel biomarkers and new therapeutic strategies for HCC,including alpha-fetoprotein-L3,glypican-3,heat shock protein 90,dickkopf WNT signaling pathway inhibitor 1,paraoxonase 1,highly up-regulated in liver cancer.Moreover,epigenetic regulation,signal pathway,cellular and molecular targets for the immunotherapy,tumor microenviroment and genome sequencing analysis may serve as the molecular expression signatures in clinical practice.For promising new treatment strategy of HCC,targeting molecular therapy based on the restoration of tumor suppressor genes lost and inhibition of oncogenic genes is attractive.The new clinical trials for other molecular-targeted agents,including pembrolizumab,nivolumab,tivantinib,lenvatinib,cabozantinib,and ramucirumab,are ongoing in clinic.Interestingly,anti-HBV drugs display an amazing therapy for HBV-related HCC.In future,the global determination of more biomarkers may provide new insights into the diagnosis of HCC.More importantly,the diagnostic markers should be used to trace patient's follow-up disease progression,guiding doctors to judge and prescribe drugs for status of an illness,prognosis and other processes.
基金National Nature Science Foundation of China under Grant! No. 19775011 under Grant !No.10075016China Nuclear Industry Sci
文摘Supersonic Molecular Beam Injection (SMBI) is a new fuelling method for Tokamaks and has recently been improved to enhance the flux of the beam and to make a survey of the cluster effect within the beam. There are a series of new phenomena, which implicate the interaction of the beam (including clusters) with the toroidal plasma of HL-1M Tokamak. The Ha signals from the edge show a regular variation around the torus. Around the injection port, the edge Hα signals are positive rectangular wave, which is consistent with that of the injection beam pulses. The edge electron temperature, measured with movable Langmuir probes, decreases by an order of magnitude and the density increases by an order of magnitude. Hα emission at the beam injection port, measured with CCD camera at an angle of 13.4 degrees to the SMBI line, shows many separate peaks within the contour plot. These peaks may show the strong emission produced by the interaction of the hydrogen clusters with the plasma. Hydrogen clusters may be produced in the beam according to the empirical scaling (Hagena) law of clustering onset, * = .here d is the nozzle diameter in μm, Po the stagnation pressure in mbar, To the source temperature in K, and k is a constant related to the gas species. If * > 100, clusters will be formed. In present experiment * is about 127.
基金supported by a grant from Fundacao de Amparo a Pesquisa no Estado de Sao Paulo(FAPESP)with grant number 2010-50304-8,under the supervision of Dr.Lucia Maria Almei Braz.
文摘Objective:To determine an algorithm for molecular diagnosis of visceral leishmaniasis(VL)by kinetoplast DNA(kDNA)(RV1/RV2)and internal transcriber spacer(ITS1)(LITSR/L5.8 S)polymerase chain reaction(PCR),complemented by ITS 1 PCR restriction fragment length polymorphism(RFLP),using peripheral blood or bone marrow aspirate from patients with suspected VL.Methods:Biological samples were submitted to the gold standard for the diagnosis of VL and molecular diagnosis represented by ITS 1 PCR,kDNA PCR,and ITS 1 PCR RFLP.The samples were obtained from seven groups:groupⅠ,82 samples from patients with confirmed VL;groupⅡ,16 samples from patients under treatment for VL;groupⅢ,14 samples from dogs with canine visceral leishmaniasis(CVL);groupⅣ,a pool of six experimentally infected sandflies(Lutzomya longipalpis);group V,18 samples from patients with confirmed tegumentary leishmaniasis(TL)and groupsⅥandⅦwere from control groups without VL.Results:The following gold standard and molecular examination results were obtained for each of the seven groups:groupⅠ:parasitologic and immunochromatographic tests showed a sensitivity of 76.3%(61 of 80)and 68.8%(55 of 80),respectively,and a sensitivity of 97.6%(80 of 82)and 92.7%(76 of 82)by ITS1 and kDNA PCR,respectively.After ITS1 PCR RFLP(HaeⅢ)analysis of the 80 positive samples,52.5%(42 of 80)generated three fragments of 180,70,and 50 bp,corresponding to the pattern of Leishmania infantum infantum;groupⅡ:negative for the parasitologic methods and positive for IrK39(100%,16 of 16),presented 12.5%(2 of 16)of positivity by ITS 1 PCR and 25.0%(4 of 16)by kDNA PCR;groupⅢ:positive in the parasitologic and serologic tests(100%,14 of 14),presented 85.7%(12 of 14)of positivity by ITS1 PCR and kDNA PCR.ITS1 PCR RFLP showed that 83.3%(10 of 12)of the canine samples contained parasites with profiles similar to L.infantum;groupⅣpresented amplifications by ITS1 PCR and kDNA PCR.ITS1 PCR products were analyzed by RFLP,generating a profile similar to that of L.infantum;groupⅤ:positive in the parasitologic examination(100%,18 of 18),presented 72.2%(13 of 18)of the samples by ITS1 PCR positive.A total of 69.2%(9 of 13)showed profiles corresponding to a Viannia complex by ITS1 PCR RFLP;and groupⅥand groupⅥwere negative by ITS 1 and kDNA molecular tests.Comparing the molecular results with the parasitologic and serologic diagnosis from groupⅠ,almost perfect agreement was found(κboth>0.80,P<0.001).ITS1 and RV1/RV2 PCR detected 90.2%(74 of 82)of the samples.Two samples positive by RV1/RV2 were negative by LITSR/L5.8 S,and six samples positive by LITSR/L5.8 S were negative by RV1/RV2.Therefore,these two systems complemented each other;they diagnosed 100%of the samples as belonging to the Leishmania genus.Conclusions:We suggest an algorithm for the molecular diagnosis of VL,which must consider previous parasitologic and serologic(immunochromatographic)diagnoses,and should combine kDNA and ITS1 to determine the Leishmania subgenus using RFLP as a complement method to define the L.infantum species.
文摘The existence of an inflammatory process in the heart muscle,related to a progressive worsening of myocardial function,different etiopathogenetic mechanisms concur and often overlap,thus making the diagnosis and the therapeutic approach complex.As the COVID-19 pandemic progresses,the effects of the disease on the organ systems and in particular on the cardiovascular system are becoming more and more profound.Cardiac involvement is a well-known event with a high percentage of findings in the heart’s magnetic field,even in asymptomatic areas.There are numerous uncertainties regarding their evolution,in the long and short term,due not only to a difficult to determine the varied clinical expression and the rarely performed intramyocardial biopsy which additionally presents diagnostic problems but also in part to different clinical prognosis.Today,the new SARS-CoV-2 virus that uses the angiotensin converting enzyme 2(ACE2)which is present at high levels in myocardial cells as its entrance it can create even severe heart injury.The pathophysiology in all of these cases can involve multiple immune and non-immune mechanisms within organs and vessels and can be occur in the clinical phases.Possible mechanisms of direct and indirect myocardial infarction in patients with COVID-19 include additional lesion and oxygen-rich and generalized inflammation response with myocardial immune hyperactivity(myocarditis).Therefore,these can occur through the excessive release of cytokines,the presence of thrombocytopenia,endocrine damage,heart failure,arrhythmias and more.Patients can show average signs of myocardial damage,and some develop spontaneous cardiac complications,such as heart failure,arrhythmias and,rarely,rare cardiogenic disorders.Pathophysiology in all of these may involve multiple mechanisms within the cytokine cephalic membrane,endocrine damage and thrombogenicity.The diagnosis of this myocardial injuri is mainly based on the myocardial enzyme troponin.This viewpoint paper explains today’s knowledge on viral myocarditis,in particular that from SARS-CoV-2 infection,if there is a connection with other possible biomolecular pathogenetic factors that can influence its natural course.In fact,it is for this reason that the pathogenetic mechanisms are analyzed and described.At the same time,its possible interaction with other parameters that are documented risk factors for cardiovascular disease was examined.Although these biomolecular findings were mainly related to necrotic parts of the myocardium,it is important to recognize that myocardial damage early for a better approach and prognosis.
文摘<b><span style="font-family:Verdana;">Background:</span></b><span style="font-family:Verdana;"> Extrapulmonary tuberculosis (EPTB) remains difficult to diagnose because the clinical specimens to be examined are often paucibacillary</span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;"> and obtained with difficulty from inaccessible sites. </span><span style="font-family:Verdana;">An updated Xpert<sup></sup></span></span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"><sup><span style="white-space:nowrap;">®</span></sup></span></span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> MTB/RIF Ultra (Ultra) test has been designed and licensed to improve sensitivity in the detection of Mycobacterium tuberculosis complex.</span></span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;"> The aim of the present study is to evaluate the performance of Ultra assay for the clinical diagnosis of EPTB in </span><span style="font-family:Verdana;">a low tuberculosis prevalence country. </span><span style="font-family:Verdana;"><b></b></span><b><b><span style="font-family:Verdana;">Methods:</span></b><span style="font-family:Verdana;"></span></b></span><span style="font-family:Verdana;"> A retrospective analysis was performed at “A.</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">O dei Colli” of Naples on consecutive extrapulmonary specimens for EPTB across a three-year period. All different types of extrapulmonary specimens were tested for EPTB by smear microscopy, culture and Ultra assay in accordance with relevant guidelines. </span><span style="font-family:Verdana;"><b></b></span><b><b><span style="font-family:Verdana;">Results:</span></b><span style="font-family:Verdana;"></span></b></span><b> </b><span style="font-family:Verdana;">A total of 606 EPTB samples, 561 culture negative EPTB and 45 culture positive EPTB were included. Using culture as reference standard, the overall sensitivities and specificities of Ultra assay were 95.6% (95% CI 84.8</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">-</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">99.5) and 97.5% (95% CI 95.8</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">-</span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">98.6) respectively. Sensitivity and specificity of Ultra for individual category of specimens w</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">ere</span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;"> also</span><span style="color:red;"> </span><span style="font-family:Verdana;">performed. </span><span style="font-family:Verdana;"><b></b></span><b><b><span style="font-family:Verdana;">Conclusion:</span></b><span style="font-family:Verdana;"></span></b></span><span style="font-family:Verdana;"> In a </span><span style="font-family:Verdana;">low-tuberculosis prevalence setting, Ultra assay confirms to have a good performance in the diagnosis of EPTB for all different extrapulmonary samples.</span></span></span>
基金funded by the International Foundation for Science(IFS),Karlavagen 108,5th floor,SE-115 26 Stockholm,Sweden(Fellow ship No.AB/21683R).
文摘Background: African animal trypanosomiasis (AAT) is caused mainly by Trypanosoma congolense, T. vivax, and T. brucei brucei and is the major constraint for livestock productivity in Sub-Saharan African countries. Information about animal trypanosomiasis status in Ivory Coast is missing, especially regarding molecular epidemiology. Therefore, this study intended to apply molecular tools to identify and characterize trypanosomes in Ivory Coast for sustainable control. Methods: 363 cattle blood samples were collected from Ferkessedougou Region in northern Ivory Coast in 2012. Buffy coat technique (BCT) and species-specific PCR assays were used to detect trypanosome species. Results: Out of 363 cattle examined with BCT, 33 were found positive with all trypanosomes species accounting for an average of 9.09% prevalence whereas polymerase chain reaction (PCR) using species-specific primers showed that 81 out of 363 cattle were infected with trypanosomes with an overall prevalence of 22.31%. Trypanosoma congolense savanah type, T. Vivax and T. brucei sl. accounted for 28.39%, 49.38% and 23.45% of the infection rate respectively. No infection with T. congo forest?type was detected. T. vivax infection was the most prevalence in the area investigated compared to the two other trypanosome species. Mixed infections with different trypanosomes species were detected accounting for 7.32% of prevalence. Regarding sexrelated prevalence, male cattles were slightly more infected than female but the difference was not significant. Conclusion: Our results showed that there was a high prevalence of AAT in livestock in Ferkessedougou Area. There is therefore a need to strengthen control policies and institute measures that help prevent the spread of the parasites for sustainable control of animal trypanosome in this area.
文摘Haemochromatosis is a genetic disease caused by hepcidin deficiency,responsible for an increase in intestinal iron absorption.Haemochromatosis is associated with homozygosity for the HFE p.Cys282Tyr mutation.However,rare cases of haemochromatosis(non-HFE haemochromatosis)can also be caused by path-ogenic variants in other genes(such as HJV,HAMP,TFR2 and SLC40A1).A working group of the International Society for the Study of Iron in Biology and Medicine(BIOIRON Society)has concluded that the classification based in different molecular subtypes is difficult to be adopted in clinical practice and has proposed a new classification approaching clinical questions and molecular complexity.The aim of the present review is to provide an update on classification,pathophysiology and therapeutic recommendations.
基金supported by the National Natural Science Foundation of China(Grant No.:81703466)the Outstanding Talents Research Start-up Fund of Xuzhou Medical University,China(Grant No.:RC20552107)Xuzhou Science and Technology Bureau,China(Grant No.:KC21292).
文摘Fluorescence imaging is a noninvasive and dynamic real-time imaging technique;however,it exhibits poor spatial resolution in centimeter-deep tissues because biological tissues are highly scattering media for optical radiation.The recently developed ultrasound-controlled fluorescence(UCF)imaging is a novel imaging technique that can overcome this bottleneck.Previous studies suggest that the effective contrast agent and sensitive imaging system are the two pivotal factors for generating high-resolution UCF images ex vivo and/or in vivo.Here,this review highlights the recent advances(2015e2020)in the design and synthesis of contrast agents and the improvement of imaging systems to realize high-resolution UCF imaging of deep tissues.The imaging performances of various UCF systems,including the signal-to-noise ratio,imaging resolution,and imaging depth,are specifically discussed.In addition,the challenges and prospects are highlighted.With continuously increasing research interest in this field and emerging multidisciplinary applications,UCF imaging with higher spatial resolution and larger imaging depth may be developed shortly,which is expected to have a far-reaching impact on disease surveillance and/or therapy.
基金supported by AESI-ISC Ⅲ grant number PI14C Ⅲ/00014
文摘Objective:To develop a new technique for diagnosis of Plasmodium knowlesi and at the same time to be able to discriminate among the diverse species of Plasmodium causing human malaria.Methods:In this study the nested multiplex malaria PCR was redesigned,targeting the 18S rR NA gene,to identify the fifth human Plasmodium species,Plasmodium knowlesi,together with the other human Plasmodium(Plasmodium falciparum,Plasmodium vivax,Plasmodium ovale and Plasmodium malariae)by amplified fragment size using only two amplification processes and including an internal reaction control to avoid false negatives.Results:The technique was validated with 91 clinical samples obtained from patients with malaria compatible symptoms.The technique showed high sensitivity(100%)and specificity(96%)when it was compared to the reference method employed for malaria diagnosis in the Instituto de Salud Carlos栿and a published real-time PCR malaria assay.Conclusions:The technique designed is an economical,sensitive and specific alternative to current diagnosis methods.Furthermore,the method might be tested in knowlesi-malaria endemic areas with a higher number of samples to confirm the quality of the method.
文摘Objective: The goal of this study was to determine the prevalence of C. trachomatis in women diagnosed with infertility attending the Outpatient Clinic of Infertility from Botucatu Medical School, UNESP, Brazil. Patients and Methods: This molecular study enrolled a total of 112 women. Among these patients, 62 presented primary infertility while 50 presented secondary infertility. The criteria for eligibility included women who were: reproductive-aged;no prior report of seroconversion for HIV;no antibiotic or vaginal cream used in the preceding 30 days;and abstinence from sexual intercourse for 72 hours before the visit. The women were submitted to a gynecological examination and cervical samples were collected with an endocervical cytobrush for molecular analysis of C. trachomatis. Results: The prevalence of chlamydial infection was 8% with similar prevalence between primary (8.1%) and secondary (8.0%) infertility. Conclusion: Considering the asymptomatic nature of chlamydial infection and its association with tubal factor infertility, there is a pressing need to incorporate the screening of C. trachomatis infection as part of the routine investigation for infertility. The early diagnostic by screening can minimize complications and reduce Public Health costs with Assisted Reproductive Technology.
基金Natural Science Foundation of China(No.81902484)China Postdoctoral Science Foundation(No.2020M670864)+2 种基金Medical and Health Talents Project of Jilin Province(Nos.2019SCZT003,2020SCZT096)Youth Support Project of Jilin Association for Science and Technology(No.202028)Project of Hepatobiliary and Pancreatic Disease Translational Medicine Platform Construction(No.2017F009)
文摘Hepatocellular carcinoma(HCC)is the most common type of primary hepatocellular carcinoma(PHC).Early diagnosis of HCC remains the key to improve the prognosis.In recent years,with the promotion of the concept of precision medicine and more in-depth analysis of the biological mechanism underlying HCC,new diagnostic methods,including emerging serum markers,liquid biopsies,molecular diagnosis,and advances in imaging(novel contrast agents and radiomics),have emerged one after another.Herein,we reviewed and analyzed scientific advances in the early diagnosis of HCC and discussed their application and shortcomings.This review aimed to provide a reference for scientific research and clinical practice of HCC.
基金This paper was partially supported by the National Institutes of Health(grants Nos.R01AI113725 and R01HD090927).
文摘Diagnosis of tuberculosis can be difficult as advances in molecular diagnosis approaches(especially nanoparticles combined with high-throughput mass spectrometry for detecting mycobacteria peptide)and personalized medicine result in many changes to the diagnostic framework.This review will address issues concerning novel technologies from bench to bed and new strategies for personalized tuberculosis diagnosis.
基金supported by the Ministry of Science and Technology(2020YFC0846300)National Science and Technology Major Project(2017ZX10103011,2018ZX10711001)+1 种基金Shenzhen Science and Technology Research and Development Project(202002073000001)China Postdoctoral Science Foundation(2019T120147,2019M660836)。
文摘The worldwide epidemic of coronavirus disease 2019(COVID-19)is ongoing.Rapid and accurate detection of the causative virus SARSCoV-2 is vital for the treatment and control of COVID-19.In this study,the comparative sensitivity of different respiratory specimen types were retrospectively analyzed using 3,552 clinical samples from 410 COVID-19 patients confirmed by Guangdong CDC(Center for Disease Control and Prevention).Except for bronchoalveolar lavage fluid(BALF),the sputum possessed the highest positive rate(73.4%–87.5%),followed by nasal swabs(53.1%–85.3%)for both severe and mild cases during the first 14 days after illness onset(d.a.o.).Viral RNA could be detected in all BALF samples collected from the severe group within 14 d.a.o.and lasted up to 46 d.a.o.Moreover,although viral RNA was negative in the upper respiratory samples,it was also positive in BALF samples in most cases from the severe group during treatment.Notably,no viral RNA was detected in BALF samples from the mild group.Despite typical ground-glass opacity observed via computed tomographic scans,no viral RNA was detected in the first three or all upper respiratory tract specimens from some COVID-19 patients.In conclusion,sputum is most sensitive for routine laboratory diagnosis of COVID-19,followed by nasal swabs.Detection of viral RNA in BALF improves diagnostic accuracy in severe COVID-19 patients.
基金The study was funded by the grant from National Key Clinical Specialist Construction Programs of China-Neonatology(Grant No.2011-873).The funding agency had no role in study design,data collection and analysis,or preparation of the manuscript.
文摘Many seizures in neonates are due to early-onset epilepsy,which is often difficult to diagnose,especially to explore the causes.Recently,the development of next-generation sequencing(NGS)has led to the discovery of a large number of genes involved in epilepsy.This may improve prompt detection of early-onset epilepsy in neonates.This study aimed at analyzing the genotype-phenotype correlations in neonates with seizures in a bid to improve the understanding of genetic diagnosis of early-onset epilepsy.Clinical features and prognosis of 15 children who underwent genetic testing having had unexplained seizures from February 2016 to May 2018 in Children’s Hospital of Chongqing Medical University were analyzed retrospectively.The salient findings were:poor response to stimulus and abnormal electroencephalogram(EEG)in the initial period were observed in the group with concomitant genetic abnormalities.Despite the recent progress in genetic technology,molecular diagnosis for neonatal-onset epilepsy can be challenging due to genetic and phenotypic heterogeneities.However,some genotypes are associated with specific clinical manifestations and EEG patterns.Therefore,in-depth understanding of genotype-phenotype correlations would be useful to clinicians managing neonates with early-onset seizures.
基金supported by the National Natural Science Foundation of China(Nos.81672090 and 81871717)
文摘Congenital heart disease(CHD)is the most common birth defect worldwide.In recent years,the widespread application of innovative molecular diagnostic technologies in clinical scenarios has obviously increased the molecular diagnostic yields of CHD,providing evidence-based guidance for medical decision-making.These molecular diagnostic technologies include chromosome microarray analysis,targeted sequencing,exome sequencing,and genome sequencing.Furthermore,high-throughput sequencing technology has performed excellently in the clinical molecular diagnosis of CHD.This review provides an overview of the current technology and applications in the molecular diagnosis of CHD.The unmet issues and future directions in adapting novel genomic testing technologies to the molecular diagnosis of CHD in clinical settings are also addressed.
基金supported by grants from Shanghai Outstanding Academic Leaders(20XD1433300)Medical-Engineering Cross Foundation of Shanghai Jiao Tong University(YG2021ZD20)+3 种基金Shuguang Project(21SG11),Innovative Research Team of High-level Local Universities in Shanghai(SHSMU-ZDCX20212700)Shanghai Research Center for Endocrine,Metabolic Diseases(2022ZZ01002)Shanghai Sixth People’s Hospital Grant(ynhg202204)Shanghai Municipal Key Clinical Specialty.
文摘A small fraction of patients diagnosed with obesity or diabetes mellitus has an underlying monogenic cause.Here,we constructed a targeted gene panel consisting of 83 genes reported to be causative for monogenic obesity or diabetes.We performed this panel in 481 patients to detect causative variants and compared these results with whole-exome sequencing(WES)data available for 146 of these patients.The coverage of targeted gene panel sequencing was significantly higher than that of WES.The diagnostic yield in patients sequenced by the panel was 32.9%with subsequent WES leading to three additional diagnoses with two novel genes.In total,178 variants in 83 genes were detected in 146 patients by targeted sequencing.Three of the 178 variants were missed by WES,although the WES-only approach had a similar diagnostic yield.For the 335 samples only receiving targeted sequencing,the diagnostic yield was 32.2%.In conclusion,taking into account the lower costs,shorter turnaround time,and higher quality of data,targeted sequencing is a more effective screening method for monogenic obesity and diabetes compared to WES.Therefore,this approach could be routinely established and used as a first-tier test in clinical practice for specific patients.
