CAJ | 학술논문

目的建立间接酶联免疫吸附法(ELISA)检测人博卡病毒(HBoV)抗体,探讨其临床应用的可行性.方法以重组表达的HBoV融合蛋白VP2作为包被抗原,建立检测HBoV抗体的间接ELISA,优化该检测方法的最佳条件,观察其精密度、敏感度和特异性,并与荧光定量PCR方法对比检测40份临床样本,同时采用免疫印迹法(Western blot)确认其符合率.结果所建立的间接ELISA最佳抗原包被浓度为2 mg/L,酶标二抗工作浓度为1∶5000,血清标本最佳稀释度为1∶200.该方法平均批内变异系数为6.87%,平均批间变异系数为4.67%.40份临床样本间接ELISA检测结果与荧光定量PCR及免疫印迹结果一致,符合率100%.结论利用VP2融合蛋白作为抗原,建立的检测血清HBoV抗体间接ELISA方法特异性强、重复性好,可用于HBoV抗体的定量和定性检测.
목적 건립간접매련면역흡부법(ELISA)검측인박잡병독(HBoV)항체,탐토기림상응용적가행성.방법 이중조표체적HBoV융합단백VP2작위포피항원,건립검측HBoV항체적간접ELISA,우화해검측방법적최가조건,관찰기정밀도、민감도화특이성,병여형광정량PCR방법대비검측40빈림상양본,동시채용면역인적법(Western blot)학인기부합솔.결과 소건립적간접ELISA최가항원포피농도위2 mg/L,매표이항공작농도위1∶5000,혈청표본최가희석도위1∶200.해방법평균비내변이계수위6.87%,평균비간변이계수위4.67%.40빈림상양본간접ELISA검측결과여형광정량PCR급면역인적결과일치,부합솔100%.결론 이용VP2융합단백작위항원,건립적검측혈청HBoV항체간접ELISA방법특이성강、중복성호,가용우HBoV항체적정량화정성검측.
Objective To establish indirect ELISA for detection of HBoV antibody and to evaluate the feasibility of its clinical use. Methods Using the HBoV fusion protein VP2 generated by recombinant expression as envelope antigen, indirect ELISA method for HBoV antibody detection was established. The optimal condition of the test was set, and precision, sensitivity and specificity were examined. The findings of the test were compared with those using quantitative fluorescence PCR in 40 clinical samples, and were validated by Western blot to evaluate the consistency between each other. Results The optimal 4.67%. The findings of this test in 40 clinical samples were 100% consistent with those by quantitative fluorescence PCR and Western blot. Conclusion The new indirect ELISA method for HBoV antibody detection using HBoV fusion protein VP2 as envelope antigen is highly specific, reproducible and useful for quantitative or qualitative detection of HBoV antibody.