Objective To develop a rapid,highly sensitive quantitative method for detecting P24 antigen based on near-infrared fluorescent microsphere immunochromatography.Methods First,we prepared a lateral flow assay test strip...Objective To develop a rapid,highly sensitive quantitative method for detecting P24 antigen based on near-infrared fluorescent microsphere immunochromatography.Methods First,we prepared a lateral flow assay test strip,and labeled the detection antibody using a fluorescent microsphere.Second,we optimized the antibody labeling conditions.Third,we optimized the detection conditions.Fourth,we created a working curve.Fifth,we conducted a methodological assessment of the established fluorescent microsphere immunochromatography method.Sixty-six clinical samples were tested,and we compared the established fluorescent microsphere immunochromatography with the quantitative ELISA method.Results According to the working curve,the detection limit of the method is 3.4 pg/mL,and the detection range is 3.4 pg/mL to 10 ng/mL.The average intra-assay recovery was 99.6%,and the Coefficient of Variation(CV)was 5.4%–8.6%;the average inter-assay recovery was 97.3%,and the CV was 8.5%–11%.The detection rate of fluorescent microsphere immunochromatography was higher than ELISA method,and had a good correlation with ELISA.Conclusion The P24 antigen quantitative detection method based on near-infrared fluorescent microsphere immunochromatography has the advantages of rapid detection,high sensitivity,and wide detection range;thus,it is suitable for early clinical diagnosis and continuous monitoring of AIDS.展开更多
目的对人类免疫缺陷病毒1型(H IV-1)p24抗原酶联免疫诊断试剂进行初步评价。方法用实时荧光PCR、EL ISA、W estern B lotting等方法检测来源于中国不同地区的124份血浆样品的H IV RNA、H IV p24、抗-H IV抗体,并对部分样品进行H IV基因...目的对人类免疫缺陷病毒1型(H IV-1)p24抗原酶联免疫诊断试剂进行初步评价。方法用实时荧光PCR、EL ISA、W estern B lotting等方法检测来源于中国不同地区的124份血浆样品的H IV RNA、H IV p24、抗-H IV抗体,并对部分样品进行H IV基因分型。结果9例H IV抗体阴性样品中有1例H IV p24抗原阳性,证明为窗口期感染;9例H IV抗体不确定者中有2例为H IV p24抗原阳性;103例H IV抗体确证为阳性者中有8例为p24抗原阳性;3例H IV抗体酶联免疫检测为阳性但确证试剂检测为阴性的样品中,均无p24抗原阳性者。结论H IV p24抗原检测可以检出H IV感染的窗口期,且可以辅助H IV抗体检测试剂对H IV感染的诊断。展开更多
We report a p24(HIV disease biomarker)detection assay using an MgO-based magnetic tunnel junction(MTJ)sensor and 20-nm magnetic nanoparticles.The MTJ array sensor with sensing area of 890×890μ2 possessing a sens...We report a p24(HIV disease biomarker)detection assay using an MgO-based magnetic tunnel junction(MTJ)sensor and 20-nm magnetic nanoparticles.The MTJ array sensor with sensing area of 890×890μ2 possessing a sensitivity of 1.39%/Oe was used to detect p24 antigens.It is demonstrated that the p24 antigens could be detected at a concentration of 0.01μg/ml.The development of bio-detection systems based on magnetic tunnel junction sensors with high-sensitivity will greatly benefit the early diagnosis of HIV.展开更多
基金supported by the National Natural Science Foundation of China[no.21677006]National Key Research and Development Project[2017YFC1200500].
文摘Objective To develop a rapid,highly sensitive quantitative method for detecting P24 antigen based on near-infrared fluorescent microsphere immunochromatography.Methods First,we prepared a lateral flow assay test strip,and labeled the detection antibody using a fluorescent microsphere.Second,we optimized the antibody labeling conditions.Third,we optimized the detection conditions.Fourth,we created a working curve.Fifth,we conducted a methodological assessment of the established fluorescent microsphere immunochromatography method.Sixty-six clinical samples were tested,and we compared the established fluorescent microsphere immunochromatography with the quantitative ELISA method.Results According to the working curve,the detection limit of the method is 3.4 pg/mL,and the detection range is 3.4 pg/mL to 10 ng/mL.The average intra-assay recovery was 99.6%,and the Coefficient of Variation(CV)was 5.4%–8.6%;the average inter-assay recovery was 97.3%,and the CV was 8.5%–11%.The detection rate of fluorescent microsphere immunochromatography was higher than ELISA method,and had a good correlation with ELISA.Conclusion The P24 antigen quantitative detection method based on near-infrared fluorescent microsphere immunochromatography has the advantages of rapid detection,high sensitivity,and wide detection range;thus,it is suitable for early clinical diagnosis and continuous monitoring of AIDS.
文摘目的对人类免疫缺陷病毒1型(H IV-1)p24抗原酶联免疫诊断试剂进行初步评价。方法用实时荧光PCR、EL ISA、W estern B lotting等方法检测来源于中国不同地区的124份血浆样品的H IV RNA、H IV p24、抗-H IV抗体,并对部分样品进行H IV基因分型。结果9例H IV抗体阴性样品中有1例H IV p24抗原阳性,证明为窗口期感染;9例H IV抗体不确定者中有2例为H IV p24抗原阳性;103例H IV抗体确证为阳性者中有8例为p24抗原阳性;3例H IV抗体酶联免疫检测为阳性但确证试剂检测为阴性的样品中,均无p24抗原阳性者。结论H IV p24抗原检测可以检出H IV感染的窗口期,且可以辅助H IV抗体检测试剂对H IV感染的诊断。
基金President’s Fund of CUHKSZ,Longgang Key Laboratory of Applied Spintronics,at The Chinese University of Hong Kong,the National Natural Science Foundation of China(Grant Nos.11974298 and 61961136006)the Shenzhen Fundamental Research Fund,China(Grant No.JCYJ20170410171958839)Shenzhen Peacock Group Plan,China(Grant No.KQTD20180413181702403).
文摘We report a p24(HIV disease biomarker)detection assay using an MgO-based magnetic tunnel junction(MTJ)sensor and 20-nm magnetic nanoparticles.The MTJ array sensor with sensing area of 890×890μ2 possessing a sensitivity of 1.39%/Oe was used to detect p24 antigens.It is demonstrated that the p24 antigens could be detected at a concentration of 0.01μg/ml.The development of bio-detection systems based on magnetic tunnel junction sensors with high-sensitivity will greatly benefit the early diagnosis of HIV.
文摘目的评价Roche电化学发光法联合检测HIV抗原抗体试剂的检测性能。方法收集包括健康献血员、人类免疫缺陷病毒(HIV)感染者、丙型肝炎病毒(HCV)抗体阳性和高危人群在内的血清样本1 327例。购买美国BBI公司的7套HIV血清转换盘共计51份血清样本。采用Roche cobas e411(架式系统)电化学全自动免疫分析仪和相应的检测试剂、校准品和质控品以及检测参数进行测定。以Roche Elecsys HIV Combi试剂盒为评估试剂,以生物梅里埃Vironostika Uni-FormⅡplus O HIV抗体试剂盒为参比试剂。定性检测血清HIV p24抗原和HIV1/2抗体并评价检测方法分析性能、结果一致性和血清转化灵敏度。结果 Roche Elecsys HIV Combi试剂盒批内和天间不精密度良好,符合临床使用要求。血清样本检测结果和参比试剂检测结果一致性完全符合。评估试剂的血清转化灵敏度比参比试剂平均提前了5.1 d。结论 Roche Elecsys HIV Combi试剂盒具有良好的精密度和血清转换灵敏度,有利于筛查HIV早期感染,能为临床HIV早期诊断、随访监测提供有效的实验室诊断依据。
