Qualitative qPCR detected HIV-1 proviral DNA with high sensitivity and specificity in clinical samples, indicating effective early diagnosis methods.
HIV-1 proviral DNA is a critical marker for the early diagnosis of vertical transmission. The objective of the study was to evaluate the performance of a qualitative qPCR for detecting HIV-1 proviral DNA. The qPCR, TaqMan system, evaluated targeted the Integrase (IN) and Long Terminal Repeat (LTR) regions of HIV-1 and the CCR5 gene as an internal control. Aiming to increase the sensitivity of qPCR, a Nested qPCR was developed. ACH-2 cells were used to construct the standard curve and standardize the assays. The assessment of the diagnostic sensitivity and specificity of qPCR was carried out using 125 clinical samples. Nested qPCR performance was evaluated on 47/125 clinical samples. qPCR showed satisfactory efficiency for all tested targets, detection limit of 40 copies/reaction of HIV-1 proviral DNA, sensitivity of 87.2% and specificity of 100%. In Nested qPCR, the use of an additional amplification step increased the sensitivity of the assay, allowing the detection of 30 copies/reaction of HIV-1 proviral DNA. The assays evaluated demonstrated the ability to detect HIV-1 proviral DNA, with high specificity and sensitivity, applicable to a variety of patients with different levels of plasma viremia and capable of detecting a variety of HIV-1 subtypes and CRFs.
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Santos et al. (2025) studied this question.
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