实时聚合酶链反应评估 HIV 感染者中男男性行为者的肛门和生殖器样本中单纯疱疹病毒 2 的脱落情况。

Real time PCR to evaluate HSV-2 shedding from anal and genital samples among men who have sex with men, living with HIV.

机构信息

Centro de Investigación sobre Enfermedades Infecciosas, Instituto Nacional de Salud Pública, Cuernavaca, Morelos.

Centro de Investigación en Enfermedades Infecciosas, Instituto Nacional de Enfermedades Respiratorias, Mexico.

出版信息

J Med Virol. 2018 Apr;90(4):745-752. doi: 10.1002/jmv.25003. Epub 2018 Jan 18.

DOI:10.1002/jmv.25003

PMID:29236293

Abstract

This study shows the relative quantification of HSV-2 by qPCR, using the MIQE Guidelines. The reaction efficiency was evaluated, and the relative quantification used the R = 2 method. The relative quantification of HSV-2 was conducted with anal and genital samples from men who have sex with men (MSM), living with HIV. The presence of a single amplification product was validated with a dissociation curves profile and the determination of the melting temperature. The limit of detection for β-globin was determined as 3.3 × 10 ng/μL, and for HSV-2 at 6.0 × 10 ng/μL. The efficiency for β-globin was 100.2% and for HSV-2 was 106.8%. From 336 MSM, 2.1% and 3.9% individuals presented anal or genital HSV-2 shedding, respectively. The HSV-2 viral load was 9.2 RU, individuals with fewer CD4 presented higher HSV-2 viral load. The qPCR method is reproducible and has optimal reaction efficiency.

摘要

本研究采用 MIQE 指南，通过 qPCR 对 HSV-2 进行相对定量。评估了反应效率，并采用 R=2 法进行相对定量。对与 HIV 共存的男男性接触者（MSM）的肛门和生殖器样本进行 HSV-2 的相对定量。通过解链曲线分析和熔解温度的测定来验证单一扩增产物的存在。β-珠蛋白的检测限为 3.3×10ng/μL，HSV-2 的检测限为 6.0×10ng/μL。β-珠蛋白的效率为 100.2%，HSV-2 的效率为 106.8%。在 336 名 MSM 中，分别有 2.1%和 3.9%的个体存在肛门或生殖器 HSV-2 脱落。HSV-2 的病毒载量为 9.2 RU，CD4 计数较少的个体 HSV-2 病毒载量较高。qPCR 方法具有良好的重现性和最佳的反应效率。