Detection of phylogenetically diverse human immunodeficiency virus type 1 groups M and O from plasma by using highly sensitive and specific generic primers.

Published

Journal Article

The high degree of genetic diversity within human immunodeficiency virus type 1 (HIV-1), which includes two major groups, M (major) and O (outlier), and various env subtypes within group M (subtypes A to J), has made designing assays that will detect all known HIV-1 strains difficult. We have developed a generic primer set based on the conserved immunodominant region of transmembrane protein gp41 that can reliably amplify as few as 10 copies/PCR of viral DNA from near-full-length clones representing group M subtypes A to H (subtypes I and J were not available). The assay is highly sensitive in detecting plasma viral RNA from HIV-1 strains of diverse geographic origins representing different subtypes of HIV-1 group M as well as HIV-1 group O. Of the 253 group M plasma specimens (subtypes A, 68 specimens; B, 71; C, 19; D, 27; E, 23; F, 33; and G, 12), 250 (98.8%) were amplified by using the gp41 M/O primer set. More importantly, all 32 (100%) group O plasma samples were also amplified with these primers. In vitro spiking experiments further revealed that the assay could reliably detect as few as 25 copies/ml of viral RNA and gave positive signals in HIV-1-seropositive specimens with plasma copy numbers below the limits of detection by all commercially available viral load assays. In addition, analysis of five seroconversion panels indicated that the assay is highly sensitive for early detection of plasma viremia during the "window period." Thus, the highly sensitive assay will be useful for early detection of HIV-1 in clinical specimens from all known HIV-1 infections, regardless of their genotypes and geographic origins.

Full Text

Duke Authors

Cited Authors

  • Yang, C; Pieniazek, D; Owen, SM; Fridlund, C; Nkengasong, J; Mastro, TD; Rayfield, MA; Downing, R; Biryawaho, B; Tanuri, A; Zekeng, L; van der Groen, G; Gao, F; Lal, RB

Published Date

  • August 1999

Published In

Volume / Issue

  • 37 / 8

Start / End Page

  • 2581 - 2586

PubMed ID

  • 10405405

Pubmed Central ID

  • 10405405

Electronic International Standard Serial Number (EISSN)

  • 1098-660X

International Standard Serial Number (ISSN)

  • 0095-1137

Language

  • eng