Precision and repeatability are challenging issues in point of care testing(POCT) analysis. Herein, we proposed a lateral flow assay(LFA) based on internal quality control microspheres to realize the accurate diagnosi...Precision and repeatability are challenging issues in point of care testing(POCT) analysis. Herein, we proposed a lateral flow assay(LFA) based on internal quality control microspheres to realize the accurate diagnosis of HbAlc in human body. Fluorescein cy5 decorated microspheres are used as labels for HbAlc detection, and BSA-fluorescein isothiocyanate(FITC) decorated microspheres are used as internal quality control labels. One test line was employed in the strip for the detection of glycosylated hemoglobin(HbAlc). This method can eliminate the interference of environmental factors(temperature, humidity,etc.) to LFA in the process of chromatography, and improve the precision and accuracy of HbAlc detection.The CV for detection of low concentration HbAlc was 1.05%, and the CV for detection of high concentration HbAlc was 0.69%. We envision the method to have great prospect in in vitro diagnosis(IVD).展开更多
This device is aimed at ensuring that the sample is uniformly and equivalently reacted with the antibody on the NC membrane in each test when the microfluidic liquid system is introduced to the chip.In this study,the ...This device is aimed at ensuring that the sample is uniformly and equivalently reacted with the antibody on the NC membrane in each test when the microfluidic liquid system is introduced to the chip.In this study,the developed microfluidic chip can avoid the presence of the sample and conjugate pads in the chip,while the precision of the chro matography system can be greatly improved using the same particles,NC membrane and antibody alongside the traditional strip.The results,taking the detection of cTnI as an example,revealed that the coefficient of variation(CV)is controlled within 4%,while the maximum record of the contrast chromatographic reagent strip can reach 15%.Additionally,the detection sensitivity can maintain the same order of magnitudes with that of the traditional chromatographic strip.With the results,the determination correlation of the developed microfluidic chip has been greatly improved.In addition,the CV of the chip in this study is greatly improved in comparison with that of the traditional strip.The biggest improvement lies in the mixing between the sample and the microspheres,indicating that this is a new approach to improve the CV of the traditional strip.展开更多
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
Nucleic acid extraction is one of the bases in molecular biology and it is the indispensable process for nucleic acid detection. Due to its excellent characteristics, the method based on magnetic beads has become one ...Nucleic acid extraction is one of the bases in molecular biology and it is the indispensable process for nucleic acid detection. Due to its excellent characteristics, the method based on magnetic beads has become one of the main methods for nucleic acid extraction.In this research, we have developed a rapid high-quality universal nucleic acid extraction kit based on magnetic beads(MBs) and named it MB-100. Some factors affecting nucleic acid extraction were optimized, such as particle size of MBs, surface groups of MBs and lysis time. Results demonstrated that micron-grade MBs@SiO_2 particles were much more appropriate for nucleic acid extraction and lysis time was 5 min. Simultaneously, the performance of MB-100 was explored. Results showed that it had much higher extraction efficiency than other two commercial kits, and the following PCR reaction also had much higher amplification efficiency.展开更多
基金supported by the State key Basic Research Program of the PRC(No.2014CB744501)the National Key Research and Development Program of China(No.2017YFA0205301)+1 种基金the National Natural Science Foundation of China(Nos.61527806,61471168 and 61871180)open Funding of State Key Laboratory of Oral Diseases(No.SKLOD20180F02)
文摘Precision and repeatability are challenging issues in point of care testing(POCT) analysis. Herein, we proposed a lateral flow assay(LFA) based on internal quality control microspheres to realize the accurate diagnosis of HbAlc in human body. Fluorescein cy5 decorated microspheres are used as labels for HbAlc detection, and BSA-fluorescein isothiocyanate(FITC) decorated microspheres are used as internal quality control labels. One test line was employed in the strip for the detection of glycosylated hemoglobin(HbAlc). This method can eliminate the interference of environmental factors(temperature, humidity,etc.) to LFA in the process of chromatography, and improve the precision and accuracy of HbAlc detection.The CV for detection of low concentration HbAlc was 1.05%, and the CV for detection of high concentration HbAlc was 0.69%. We envision the method to have great prospect in in vitro diagnosis(IVD).
基金financially supported by National Natural Science Foundation of China(Nos.81902153,61871180,62071119 and 61971187)Jiangsu Provincial Key Research and Development Program(No.BE 2018695)。
文摘This device is aimed at ensuring that the sample is uniformly and equivalently reacted with the antibody on the NC membrane in each test when the microfluidic liquid system is introduced to the chip.In this study,the developed microfluidic chip can avoid the presence of the sample and conjugate pads in the chip,while the precision of the chro matography system can be greatly improved using the same particles,NC membrane and antibody alongside the traditional strip.The results,taking the detection of cTnI as an example,revealed that the coefficient of variation(CV)is controlled within 4%,while the maximum record of the contrast chromatographic reagent strip can reach 15%.Additionally,the detection sensitivity can maintain the same order of magnitudes with that of the traditional chromatographic strip.With the results,the determination correlation of the developed microfluidic chip has been greatly improved.In addition,the CV of the chip in this study is greatly improved in comparison with that of the traditional strip.The biggest improvement lies in the mixing between the sample and the microspheres,indicating that this is a new approach to improve the CV of the traditional strip.
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 Natural Science Foundation of China (61527806, 61471168, 61571187)China Post-doctoral Science Foundation (2016T90403)+1 种基金the Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province [(2013) 448]the Project of Scientific Innovation Team of Ningbo (2015B11050)
文摘Nucleic acid extraction is one of the bases in molecular biology and it is the indispensable process for nucleic acid detection. Due to its excellent characteristics, the method based on magnetic beads has become one of the main methods for nucleic acid extraction.In this research, we have developed a rapid high-quality universal nucleic acid extraction kit based on magnetic beads(MBs) and named it MB-100. Some factors affecting nucleic acid extraction were optimized, such as particle size of MBs, surface groups of MBs and lysis time. Results demonstrated that micron-grade MBs@SiO_2 particles were much more appropriate for nucleic acid extraction and lysis time was 5 min. Simultaneously, the performance of MB-100 was explored. Results showed that it had much higher extraction efficiency than other two commercial kits, and the following PCR reaction also had much higher amplification efficiency.
