The development, evaluation, performance and validation of micro-PCR and extractor for the quantification of HIV-1 &-2 RNA

HIV infection has been a global public health threat and reported ~ 40 million deaths. AIDS is caused by the retrovirus, HIV-1, which is transmitted via body fluids and secretions during the time AIDS in HIV-2 infection is twice as compared to an HIV-1. Q-PCR is the only available method that requires trained personnel and lab infrastructure. Micro-PCR is a portable Q-PCR developed by Bigtec Bangalore, India, which is simple, accurate, fast, and operationalized in remote places where diagnostic services are not easily accessible in the country. This micro-PCR can determine simultaneously HIV-1 and HIV-2 viral load using a TruePrep™ extractor device for RNA isolation. 5.0 ml blood samples were collected at the blood collection centre at AIIMS, New Delhi, India. Samples were screened for serology and a comparison of HIV-1/-2 RNA was done between QPCR and micro-PCR in the samples. The micro-PCR assay of HIV-RNA has compared well with those from real-time PCR (r = 0.99, p > 0.002). Micro-PCR has good inter and intra-assay reproducibility over a wide dynamic range (1.0x10 2 to 1.0x10 8 IU/ml). The linear dynamic range was 10 2 to 10 8 . The clinical and analytical specificity of the assay was comparable i.e., 100%. Intra-assay and inter-assay coefficient of variation ranged from 1.17–3.15% and from 0.02–0.46% respectively. This micro-PCR is reliable, accurate and reproducible for early HIV-1 and HIV-2 viral load detection. This can easily be used in the field and in remote places where quantification of both HIV-1/-2 is not reachable.