中国麻风皮肤病杂志 ›› 2020, Vol. 36 ›› Issue (9): 528-532.doi: 10.12144/zgmfskin202009528

• 论著 • 上一篇    下一篇

重组聚合酶扩增法在HIV-1 DNA检测中的初步应用

董潇潇,王燕,董晓庆,许文炯,吴咏梅,张洪英   

  1. 南京市疾病预防控制中心,江苏南京,210003
  • 出版日期:2020-09-15 发布日期:2020-08-21
  • 通讯作者: 张洪英， E-mail: 504937000@qq.com

Preliminary application of recombinase polymerase amplification in detecting HIV-1 DNA

DONG Xiaoxiao, WANG Yan, DONG Xiaoqing, XU Wenjiong, WU Yongmei, ZHANG Hongying   

  1. Nanjing Municipal Center for Disease Control and Prevention, Nanjing 210003, China
  • Online:2020-09-15 Published:2020-08-21
  • Contact: ZHANG Hongying, E-mail: 504937000@qq.com

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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。

关键词: 艾滋病, 重组聚合酶扩增, 等温扩增技术

Abstract: 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. 

Key words: AIDS, recombinase polymerase amplification, isothermal amplification