Background: The detection of acute HIV infection (AHI) among high risk
populations can help reduce secondary transmission of HIV. The nucleic acid testing
(NAT) can shorten the test window period by up to 7-12 days. In this study, we
describe an in-house NAT based on the multiplex nested RT-PCR method to detect the
HIV RNA. We also evaluated it in a high risk cohort in Beijing.
Methods: Four primer pairs were designed and evaluated for the detection of different HIV-1 subtypes in
group M. Multiplex RT-PCR and nested PCR were performed. The sensitivity, specialty, primers compatibility among
HIV subtypes were evaluated simultaneously. In an MSM cohort in Beijing during a 3-year period, a total of 11,808 blood
samples that were negative by ELISA or indeterminate by Western blot were analyzed by this multiplex nested RT-PCR
with pooling strategy.
Results: The multiplex nested RT-PCR was successfully applied for the detection of at least six HIV-1 subtypes. The
sensitivity was 40 copies/ml and the specificity was 100%. A total of 29 people were tested HIV-1 positive with acute
infection in a MSM cohort of Beijing during a 3 years period.
Conclusion: This multiplex nested RT-PCR provides a useful tool for the rapid detection of acute HIV-1 infection. When
used in combination with the 3rd generation ELISA, it can improve the detection rate of HIV infection, especially in the
source limited regions.