Chemiluminescence immunoassay(CLⅠA) has always been a great challenge in detecting cardiac troponin Ⅰ(c Tn Ⅰ) in whole blood samples without centrifugation because of the interference of red blood cells and low sen...Chemiluminescence immunoassay(CLⅠA) has always been a great challenge in detecting cardiac troponin Ⅰ(c Tn Ⅰ) in whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Ⅰn this study, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotinconjugated c Tn Ⅰ antibody and detected by streptavidin/acridine aster-conjugated polychloromethylstyrene microspheres(PCMS). After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the c Tn Ⅰ concentrations of the serum samples,plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62%~5.67%. The assay was linear over the studied range of 0.01-50.00 ng/mL, and no hook effect was found when c Tn Ⅰ concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit(Abbott assay kit), the correlation coefficient was 0.9859. A washing-free CLⅠA was established for the rapid detection of c Tn Ⅰ in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and PCMS for signal amplification, which showed great potential in clinical application.展开更多
A chemiluminescence enzyme immunoassay based on magnetic microparticles (MmPs-CLEIA) was developed to evaluate serum a-fetoprotein (AFP) in parallel with traditional colorimetric enzyme-linked immunosorbent assay (ELI...A chemiluminescence enzyme immunoassay based on magnetic microparticles (MmPs-CLEIA) was developed to evaluate serum a-fetoprotein (AFP) in parallel with traditional colorimetric enzyme-linked immunosorbent assay (ELISA).A systematic comparison between the MmPs-CLEIA and colorimetric ELISA concluded that the MPs-CLEIA exhibited fewer dosages of immunoreagents,less total assay time,and better linearity,recovery,precision,sensitivity and validity.AFP was detected in forty human serum samples by the proposed MPs-CLEIA and ELISA,and the results were compared with commercial electrochemiluminescence immunoassay (ECLIA) kit.The correlation coefficient between MPs-CLEIA and ELISA was obtained with R 2 0.6703;however,the correlation between MPs-CLEIA and ECLIA (R 2 0.9582) was obviously better than that between colorimetric ELISA and ECLIA (R 2 0.6866).展开更多
The on-going global pandemic of coronavirus disease 2019(COVID-19)caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has been underway for about 11 months.Through November ...The on-going global pandemic of coronavirus disease 2019(COVID-19)caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has been underway for about 11 months.Through November 20,2020,51 detection kits for SARS-CoV-2 nucleic acids(24 kits),antibodies(25 kits),or antigens(2 kits)have been approved by the National Medical Products Administration of China(NMPA).Convenient and reliable SARS-CoV-2 detection assays are urgently needed worldwide for strategic control of the pandemic.In this review,the detection kits approved in China are summarised and the three types of tests,namely nucleic acid,serological and antigen detection,which are available for the detection of COVID-19 are discussed in detail.The development of novel detection kits will lay the foundation for the control and prevention of the COVID-19 pandemic globally.展开更多
The immense patient number caused by coronavirus disease 2019(COVID-19)global pandemic brings the urge for more knowledge about its immunological features,including the profile of basic immune parameters.In this study...The immense patient number caused by coronavirus disease 2019(COVID-19)global pandemic brings the urge for more knowledge about its immunological features,including the profile of basic immune parameters.In this study,eighty-eight reported COVID-19 patients in Wuhan were recruited from January to February,2020,including 32 severe/critical cases and56 mild/moderate cases.Their mean age was 56.43 years(range 17–83)and gender ratio(male/female)was 43:45.We tested SARS-CoV-2 RNA with commercial kits,investigated the level of serologic IgM and IgG antibodies against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)using magnetic particle chemiluminescence immunoassays,and compared the results of serologic tests and nucleic acid test(NAT).Among 88 patients,95.45%were confirmed as positive by the combination of NAT and antibody test,which was significantly higher(P<0.001)than by single nucleic acid test(73.86%)or serologic test(65.91%).Then the correlation between temporal profile and the level of antibody response was analyzed.It showed that seroconversion started on day 5 after disease onset and IgG level was rose earlier than IgM.Comparison between patients with different disease severity suggested early seroconversion and high antibody titer were linked with less severe clinical symptoms.These results supported the combination of serologic testing and NAT in routine COVID-19 diagnosis and provided evidence on the temporal profile of antibody response in patients with different disease severity.展开更多
基金financially supported by National Natural Science Foundation of China (Nos.81902153,61871180,62071119 and 61971187)Jiangsu Provincial Key Research and Development Program (Nos.BA2020016 and BE 2018695)。
文摘Chemiluminescence immunoassay(CLⅠA) has always been a great challenge in detecting cardiac troponin Ⅰ(c Tn Ⅰ) in whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Ⅰn this study, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotinconjugated c Tn Ⅰ antibody and detected by streptavidin/acridine aster-conjugated polychloromethylstyrene microspheres(PCMS). After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the c Tn Ⅰ concentrations of the serum samples,plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62%~5.67%. The assay was linear over the studied range of 0.01-50.00 ng/mL, and no hook effect was found when c Tn Ⅰ concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit(Abbott assay kit), the correlation coefficient was 0.9859. A washing-free CLⅠA was established for the rapid detection of c Tn Ⅰ in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and PCMS for signal amplification, which showed great potential in clinical application.
基金supported by the National Basic Research Program of China (973 Program,no. 2007CB714507)National Nature Science Foundation of China (no. 90813015)
文摘A chemiluminescence enzyme immunoassay based on magnetic microparticles (MmPs-CLEIA) was developed to evaluate serum a-fetoprotein (AFP) in parallel with traditional colorimetric enzyme-linked immunosorbent assay (ELISA).A systematic comparison between the MmPs-CLEIA and colorimetric ELISA concluded that the MPs-CLEIA exhibited fewer dosages of immunoreagents,less total assay time,and better linearity,recovery,precision,sensitivity and validity.AFP was detected in forty human serum samples by the proposed MPs-CLEIA and ELISA,and the results were compared with commercial electrochemiluminescence immunoassay (ECLIA) kit.The correlation coefficient between MPs-CLEIA and ELISA was obtained with R 2 0.6703;however,the correlation between MPs-CLEIA and ECLIA (R 2 0.9582) was obviously better than that between colorimetric ELISA and ECLIA (R 2 0.6866).
基金supported by the National Key Program for Infectious Disease of China(2018ZX10101002)the National Key Research and Development Program of China(2016YFD0500301,2020YFC0840900)。
文摘The on-going global pandemic of coronavirus disease 2019(COVID-19)caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has been underway for about 11 months.Through November 20,2020,51 detection kits for SARS-CoV-2 nucleic acids(24 kits),antibodies(25 kits),or antigens(2 kits)have been approved by the National Medical Products Administration of China(NMPA).Convenient and reliable SARS-CoV-2 detection assays are urgently needed worldwide for strategic control of the pandemic.In this review,the detection kits approved in China are summarised and the three types of tests,namely nucleic acid,serological and antigen detection,which are available for the detection of COVID-19 are discussed in detail.The development of novel detection kits will lay the foundation for the control and prevention of the COVID-19 pandemic globally.
基金supported by the Emergency Scientific Research Project for COVID-19 from Wuhan City(Grant No.2020020101010008)。
文摘The immense patient number caused by coronavirus disease 2019(COVID-19)global pandemic brings the urge for more knowledge about its immunological features,including the profile of basic immune parameters.In this study,eighty-eight reported COVID-19 patients in Wuhan were recruited from January to February,2020,including 32 severe/critical cases and56 mild/moderate cases.Their mean age was 56.43 years(range 17–83)and gender ratio(male/female)was 43:45.We tested SARS-CoV-2 RNA with commercial kits,investigated the level of serologic IgM and IgG antibodies against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)using magnetic particle chemiluminescence immunoassays,and compared the results of serologic tests and nucleic acid test(NAT).Among 88 patients,95.45%were confirmed as positive by the combination of NAT and antibody test,which was significantly higher(P<0.001)than by single nucleic acid test(73.86%)or serologic test(65.91%).Then the correlation between temporal profile and the level of antibody response was analyzed.It showed that seroconversion started on day 5 after disease onset and IgG level was rose earlier than IgM.Comparison between patients with different disease severity suggested early seroconversion and high antibody titer were linked with less severe clinical symptoms.These results supported the combination of serologic testing and NAT in routine COVID-19 diagnosis and provided evidence on the temporal profile of antibody response in patients with different disease severity.
