重组聚合酶扩增技术(RPA)定量检测淋巴细胞中的HIV-1pol基因

Quantitative detection of HIV-1 pol gene in lymphocytes by recombinase polymerase-mediatedisothermal amplification(RPA)

目的建立一种方便快捷的重组聚合酶介导等温扩增技术定量检测HIV-1 DNA的方法。方法参考文献合成1对引物,选择内参基因,用标准品扩增结果绘制标准曲线,对HIV血清学确证阳性(HIV抗体阳性)样本的淋巴细胞总DNA进行检测。结果 HIV-1 DNA标准品在10^2~10^6拷贝线性关系良好,R=0. 99,HIV血清学确证阳性病人淋巴细胞检测结果均为HIV DNA阳性。结论本研究初步建立了内标法定量快速检测淋巴细胞中HIV-1 DNA的方法,为HIV感染检测提供一个新的方法。

Objective To develop a convenient and rapid detection method for quantitative detection of HIV-1 DNA by recombinase polymerase-mediated isothermal amplification(RPA). Methods One pairs of primers were synthesized according to references,internal reference genes were selected,and a standard curve was drawn by standard gene amplification result to detect the total lymphocyte DNA of the HIV serologically positive(HIV antibody positive) sample. Results The standard of HIV-1 DNA from 10^2 copies to 10^6 copies has good linear relationship,R = 0. 99,and the lymphocyte test results of HIV-positive patients with positive serology were positive for HIV DNA. Conclusion This study initially established a method for quantitative detection of HIV-1 DNA in lymphocytes by internal standard method,and provided a new method for HIV infection detection.