CAJ | 학술논문

目的应用量子点荧光标记技术和间接免疫法,建立一种新的汉坦病毒(HV)感染的免疫检测技术.方法采用碳二亚胺交联法,在水溶性量子点表面修饰protein G和羊抗人IgG.以HV抗原片为固相载体,QDs-PG-IgG复合物为标记二抗,检测待检血清中的HV特异性抗体,并对检测条件进行优化.结果量子点与羊抗人IgG的最佳偶联条件:反应时间2h、pH 6.0、羊抗人IgG浓度20 μg/ml.检测体系的最佳反应条件:量子点标记二抗的最适工作浓度为1:200,HV感染阳性血清检测的最大稀释度为1:1280.结论成功建立了简便、快速的HV感染的量子点标记免疫检测方法,该方法具有良好的灵敏度和特异性,抗荧光猝灭性强,为进一步实现HV抗体的单分子定量检测奠定基础.
목적 응용양자점형광표기기술화간접면역법,건립일충신적한탄병독(HV)감염적면역검측기술.방법 채용탄이아알교련법,재수용성양자점표면수식protein G화양항인IgG.이HV항원편위고상재체,QDs-PG-IgG복합물위표기이항,검측대검혈청중적HV특이성항체,병대검측조건진행우화.결과 양자점여양항인IgG적최가우련조건:반응시간2h、pH 6.0、양항인IgG농도20 μg/ml.검측체계적최가반응조건:양자점표기이항적최괄공작농도위1:200,HV감염양성혈청검측적최대희석도위1:1280.결론 성공건립료간편、쾌속적HV감염적양자점표기면역검측방법,해방법구유량호적령민도화특이성,항형광졸멸성강,위진일보실현HV항체적단분자정량검측전정기출.
Objective To develop a new method to detect anti-Hantavirus IgG antibodies (HV IgG) based on quantum dots (QDs) and indirect immune technique. Methods The carbodiimide crosslinking method was used to couple protein G and goat antihuman IgG on the surface of water-solubility QDs.The coverglass covered HV antigen was used as carrier,and QDs-PG-IgG conjugates was used as labeled second antibody to detect the HV-IgG in the serum samples. The detecting conditions were optimized.Results The optimum reaction time,pH and goat antihuman IgG concentration for conjugating the QDs with goat antihuman IgG were 6.0,2h,and 20μg/ml,respectively.The optimum working dilution of QDs-PG-IgG conjugates was 1: 200.The detection limit of the serum samples was about 1:1280 dilution.Conclusion The method established in this study has been demonstrated to be a specific,sensitive,rapid test for detecting HV antibodies,laying the foundation of single molecule detection.The anti-fluorescence quenching ability of this method was significant improved.