摘要
目的:明确重组聚合酶扩增法(RPA)对HIV-1 DNA检测的敏感性和特异性。方法:合成HIV-1 pol基因并克隆至pUC57载体,命名为pHIV-1-pol,作为RPA实验的阳性对照;以无RNA酶水作为阴性对照。对13例临床初诊患者进行RPR检测,并与HIV-1抗体检测结果进行验证比较。结果:RPA方法等温扩增20 min即可有效扩增目的片段,检测浓度最低可为101 copies/μL。三份HIV Ab(-)样本及阴性对照均未产生特异性扩增,阳性对照出现了特异性扩增。RPR检测阳性结果与HIV-1抗体检测结果完全一致,患者1、2、7、10、12检测为阳性。结论:RPA方法快速简单,特异性和敏感性高。
Objective:To determine the sensitivity and specificity of recombinase polymerase amplification(RPA)in detecting HIV-1 DNA.Methods:HIV-1 pol gene was synthesized and cloned into pUC57 vector,named as pHIV-1-pol.pHIV-1-pol was the positive control and the RNase-free water was used as a negative control of RPA.Samples from 13 patients were detected with RPA and the results were compared with HIV-1 antibody detection.Results:The purpose segment successfully was amplified by RPA for 20 min and the minimum concentration plasmid DNA was 101 copies/μL.There was no amplification products in the negative control and 3 samples with HIV Ab(-).The specific amplification products were found in the positive control.The result of RPA was accordance with HIV antibody detection.Patient 1,2,7,10 and 12 was positive for RPA and HIV antibody detection.Conclusion:RPA is a fast and simple method with high sensitivity and spectivity in detecting HIV-1 DNA.
作者
董潇潇
王燕
董晓庆
许文炯
吴咏梅
张洪英
DONG Xiaoxiao;WANG Yan;DONG Xiaoqing;XU Wenjiong;WU Yongmei;ZHANG Hongying(Nanjing Municipal Center for Disease Control and Prevention,Nanjing 210003,China)
出处
《中国麻风皮肤病杂志》
2020年第9期528-532,共5页
China Journal of Leprosy and Skin Diseases
基金
江苏省预防医学科研项目(编号:Y2015002)。
关键词
艾滋病
重组聚合酶扩增
等温扩增技术
AIDS
recombinase polymerase amplification
isothermal amplification
