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

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 −ΔCq 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 −5 ng/μL, and for HSV‐2 at 6.0 × 10 −6 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.