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J Virol Methods. 2015 Mar;214:46-53. doi: 10.1016/j.jviromet.2015.01.006. Epub 2015 Feb 11.

Single genome amplification and standard bulk PCR yield HIV-1 envelope products with similar genotypic and phenotypic characteristics.

Author information

1
Department of Medicine, Boston University School of Medicine, Boston, MA, USA.
2
Boston University School of Public Health, Boston, MA, USA.
3
Department of Medicine, Boston University School of Medicine, Boston, MA, USA. Electronic address: msagar@bu.edu.

Abstract

Recent studies suggest that single genome amplification (SGA) as compared to standard bulk PCR and virus stocks from 293T transfection versus short term passage in peripheral blood mononuclear cells (PBMC) yield a less biased representation of HIV-1 envelope characteristics. In 9 different subjects, genetic diversity, divergence, and population structure were not significantly different among SGA or bulk PCR amplified envelope V1-V3 segments. In these subjects, 293T transfection derived virus stocks with SGA or bulk PCR amplified envelopes have similar infectivity, replication kinetics, co-receptor usage, and neutralization susceptibility. While PBMC passage as compared to the 293T derived virus stocks had similar co-receptor usage, PBMC viruses were less neutralization susceptible to some specific antibodies. Our results suggest that the method of envelope sequence amplification, either SGA or bulk PCR, does not have a significant impact on the genotypic and phenotypic properties of the virus envelope quasispecies.

KEYWORDS:

Diversity; Envelope; HIV-1; Neutralization; Replication; Single genome amplification

PMID:
25681527
PMCID:
PMC4394631
DOI:
10.1016/j.jviromet.2015.01.006
[Indexed for MEDLINE]
Free PMC Article

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