目的建立新型冠状病毒(2019-nCoV)、甲型流感病毒(Flu A)和乙型流感病毒(Flu B)胶体金抗原联合快速检测试剂的制备工艺,并系统评价试剂的性能指标。方法采用柠檬酸三钠两步还原法制备胶体金溶液,用抗2019-nCoV核衣壳蛋白(NP)单抗、抗Flu...目的建立新型冠状病毒(2019-nCoV)、甲型流感病毒(Flu A)和乙型流感病毒(Flu B)胶体金抗原联合快速检测试剂的制备工艺,并系统评价试剂的性能指标。方法采用柠檬酸三钠两步还原法制备胶体金溶液,用抗2019-nCoV核衣壳蛋白(NP)单抗、抗Flu A NP单抗、抗Flu B NP单抗及多肽作为标记抗体,硝酸纤维素膜上分别包被抗2019-nCoV NP单抗和抗多肽单抗制备新型冠状病毒抗原试纸条,另一硝酸纤维素膜上分别包被抗Flu A NP单抗、抗Flu B NP单抗和抗多肽单抗制备甲/乙型流感病毒抗原试纸条。对试剂盒最低检出限、重复性、交叉反应及临床诊断符合率进行性能评价。结果检测国家参考品,试剂盒最低检出限和重复性均符合要求;分析特异性研究显示,试剂盒与16种呼吸道相关病原体毒株均无交叉反应性。临床研究显示,检测新型冠状病毒总符合率为99.11%(111/112),甲型流感病毒总符合率为98.78%(81/82),乙型流感病毒诊断总符合率为97.56%(80/82)。结论新型冠状病毒和甲/乙型流感病毒抗原联合快速检测试剂检测灵敏度和特异性高,出结果快,操作便携,可用于新型冠状病毒和甲/乙型流感病毒的快速筛查及鉴别诊断。展开更多
目的以羧基化Fe3O4纳米酶为信号标记物,建立兼具可视化判读及高灵敏定量的新型纳米酶纸芯片(nanozyme-based immunochromatographic paper chip,NIPC),进行肌钙蛋白I(cTnI)的床旁快速检测。方法采用侧流免疫层析原理建立了可视化快速定...目的以羧基化Fe3O4纳米酶为信号标记物,建立兼具可视化判读及高灵敏定量的新型纳米酶纸芯片(nanozyme-based immunochromatographic paper chip,NIPC),进行肌钙蛋白I(cTnI)的床旁快速检测。方法采用侧流免疫层析原理建立了可视化快速定量检测cTnI的纳米酶免疫纸芯片,并对50例临床血清样本进行检测。结果本研究制备的肌钙蛋白I纳米酶免疫纸芯片(cTnI-NIPC)最低检测限为0.015 ng/mL,线性范围为0.03~12 ng/mL;完成检测时间为20min;临床样本测试,与临床检测结果具有良好的相关性(r=0.99,P<0.01),灵敏度为84.62%,特异性为95.83%。结论初步建立了新型肌钙蛋白I纳米酶免疫纸芯片,为临床提供了一种灵敏、快速、定性及定量检测人血清肌钙蛋白I的检测方法。展开更多
We collected blood from coronavirus disease 2019(COVID-19)convalescent individuals and investigated SARS-CoV-2-specific humoral and cellular immunity in these discharged patients.Follow-up analysis in a cohort of 171 ...We collected blood from coronavirus disease 2019(COVID-19)convalescent individuals and investigated SARS-CoV-2-specific humoral and cellular immunity in these discharged patients.Follow-up analysis in a cohort of 171 patients at 4-11 months after the onset revealed high levels of IgG antibodies.A total of 78.1%(164/210)of the specimens tested positive for neutralizing antibody(NAb).SARS-CoV-2 antigen peptide pools-stimulated-IL-2 and-IFN-y response can distinguish COVID-19 convalescent individuals from healthy donors.Interestingly,NAb survival was significantly affected by the antigen peptide pools-stimulated-IL-2 response,-IL-8 response,and-IFN-y response.The antigen peptide pools-activated CD8+T cell counts were correlated with NAb.The antigen peptide pools-activated natural killer(NK)cell counts in convalescent individuals were correlated with NAb and disease severity.Our data suggested that the development of NAb is associated with the activation of T cells and NK cells.Our work provides a basis for further analysis of the protective immunity to SARS-CoV-2 and for understanding the pathogenesis of COVID-19.It also has implications for the development of an effective vaccine for SARS-CoV-2 infection.展开更多
Background:Accurate quantification of 17-hydroxyprogesterone(17-OHP)in serum is vital for clinical and research applications.However,inter-laboratory variability in test results exists owing to the lack of a standardi...Background:Accurate quantification of 17-hydroxyprogesterone(17-OHP)in serum is vital for clinical and research applications.However,inter-laboratory variability in test results exists owing to the lack of a standardized reference measurement procedure(RMP).Therefore,in this study,we developed a highly accurate,cost-effective,and user-friendly candidate RMP(cRMP)for analyzing 17-OHP in serum.Methods:We quantified 17-OHP in serum using a one-step liquid–liquid extraction method with the addition of 17-OHP-^(13)C_(3),followed by liquid chromatography–tandem mass spectrometry.The ability of these methods to suppress interference was evaluated by chromatographic analysis.We assessed accuracy,specificity,the lower limit of quantitation,linearity,and matrix effects by following the standards specified by the Clinical and Laboratory Standards Institute.The consistency between the developed cRMP and the chemiluminescence method was evaluated through experiments with 120 clinical samples.Results:The developed cRMP required only 8 min for accurate quantification of serum 17-OHP without bias from matrix effects or interference from 19 metabolites added as potential interferents.The method exhibited favorable measurement performance,with a quantitation limit of 0.086 ng/mL,linear range of 0.1–400 ng/mL,a total imprecision of≤2.90%,spike recovery of 100.1%–100.6%,and relative deviations from assigned target values(RfB Institution)of−2.91%to 1.10%.The cRMP demonstrated good consistency with the conventional assay(chemiluminescence method),with a correlation coefficient R of 0.96977.Conclusion:A cRMP with high accuracy,cost-effectiveness,and convenient operation was developed for quantifying 17-OHP in serum.Its simplicity and robust performance make it an invaluable addition to the arsenal of analytical tools available for laboratories.This method can enhance the accuracy and reliability of 17-OHP measurements across various laboratories.展开更多
文摘目的建立新型冠状病毒(2019-nCoV)、甲型流感病毒(Flu A)和乙型流感病毒(Flu B)胶体金抗原联合快速检测试剂的制备工艺,并系统评价试剂的性能指标。方法采用柠檬酸三钠两步还原法制备胶体金溶液,用抗2019-nCoV核衣壳蛋白(NP)单抗、抗Flu A NP单抗、抗Flu B NP单抗及多肽作为标记抗体,硝酸纤维素膜上分别包被抗2019-nCoV NP单抗和抗多肽单抗制备新型冠状病毒抗原试纸条,另一硝酸纤维素膜上分别包被抗Flu A NP单抗、抗Flu B NP单抗和抗多肽单抗制备甲/乙型流感病毒抗原试纸条。对试剂盒最低检出限、重复性、交叉反应及临床诊断符合率进行性能评价。结果检测国家参考品,试剂盒最低检出限和重复性均符合要求;分析特异性研究显示,试剂盒与16种呼吸道相关病原体毒株均无交叉反应性。临床研究显示,检测新型冠状病毒总符合率为99.11%(111/112),甲型流感病毒总符合率为98.78%(81/82),乙型流感病毒诊断总符合率为97.56%(80/82)。结论新型冠状病毒和甲/乙型流感病毒抗原联合快速检测试剂检测灵敏度和特异性高,出结果快,操作便携,可用于新型冠状病毒和甲/乙型流感病毒的快速筛查及鉴别诊断。
基金This work was supported by the National Key Research and Development Program of China(2018YFE0204500)Centrally Guided Hubei Provincial Science and Technology Development Fund Project(2020ZYYD012)the Key Project for Anti-2019 Novel Coronavirus Pneumonia from the National Key Research and Development Program of China(2020YFC0845500).
文摘We collected blood from coronavirus disease 2019(COVID-19)convalescent individuals and investigated SARS-CoV-2-specific humoral and cellular immunity in these discharged patients.Follow-up analysis in a cohort of 171 patients at 4-11 months after the onset revealed high levels of IgG antibodies.A total of 78.1%(164/210)of the specimens tested positive for neutralizing antibody(NAb).SARS-CoV-2 antigen peptide pools-stimulated-IL-2 and-IFN-y response can distinguish COVID-19 convalescent individuals from healthy donors.Interestingly,NAb survival was significantly affected by the antigen peptide pools-stimulated-IL-2 response,-IL-8 response,and-IFN-y response.The antigen peptide pools-activated CD8+T cell counts were correlated with NAb.The antigen peptide pools-activated natural killer(NK)cell counts in convalescent individuals were correlated with NAb and disease severity.Our data suggested that the development of NAb is associated with the activation of T cells and NK cells.Our work provides a basis for further analysis of the protective immunity to SARS-CoV-2 and for understanding the pathogenesis of COVID-19.It also has implications for the development of an effective vaccine for SARS-CoV-2 infection.
基金Science and Technology Project of Dongguan,Grant/Award Number:20211800905072Graduate Education Innovation Program Project of Guangdong Province,Grant/Award Number:2022KCXTD010+6 种基金Discipline Construction Project of Guangdong Medical University,Grant/Award Numbers:4SG22098G,4SG22259G,4SG23030G,4SG23143GNational Natural Science Foundation of China,Grant/Award Numbers:81870016,82270013Guangdong Basic and Applied Basic Research Foundation,Grant/Award Number:2022A1515010525Guangdong Medical UniversityShenzhen YHLO Biotech Co.,Ltd.Huazhong University of Science and TechnologyShenzhen Maternal and Child Health Care Hospital。
文摘Background:Accurate quantification of 17-hydroxyprogesterone(17-OHP)in serum is vital for clinical and research applications.However,inter-laboratory variability in test results exists owing to the lack of a standardized reference measurement procedure(RMP).Therefore,in this study,we developed a highly accurate,cost-effective,and user-friendly candidate RMP(cRMP)for analyzing 17-OHP in serum.Methods:We quantified 17-OHP in serum using a one-step liquid–liquid extraction method with the addition of 17-OHP-^(13)C_(3),followed by liquid chromatography–tandem mass spectrometry.The ability of these methods to suppress interference was evaluated by chromatographic analysis.We assessed accuracy,specificity,the lower limit of quantitation,linearity,and matrix effects by following the standards specified by the Clinical and Laboratory Standards Institute.The consistency between the developed cRMP and the chemiluminescence method was evaluated through experiments with 120 clinical samples.Results:The developed cRMP required only 8 min for accurate quantification of serum 17-OHP without bias from matrix effects or interference from 19 metabolites added as potential interferents.The method exhibited favorable measurement performance,with a quantitation limit of 0.086 ng/mL,linear range of 0.1–400 ng/mL,a total imprecision of≤2.90%,spike recovery of 100.1%–100.6%,and relative deviations from assigned target values(RfB Institution)of−2.91%to 1.10%.The cRMP demonstrated good consistency with the conventional assay(chemiluminescence method),with a correlation coefficient R of 0.96977.Conclusion:A cRMP with high accuracy,cost-effectiveness,and convenient operation was developed for quantifying 17-OHP in serum.Its simplicity and robust performance make it an invaluable addition to the arsenal of analytical tools available for laboratories.This method can enhance the accuracy and reliability of 17-OHP measurements across various laboratories.
