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Recovery of APOBEC3-edited human immunodeficiency virus G->A hypermutants by differential DNA denaturation PCR.

Abstract : Virus genomes from the same family may exhibit a wide range in their DNA GC content, whereas viral hypermutants differ substantially in GC content from their parental genomes. As AT-rich DNA melts at lower temperatures than GC-rich DNA, use of a lower denaturation temperature during PCR should allow differential amplification of AT-rich genomes or variants within a quasispecies. The latter situation has been explored explicitly in a two-step process by using a series of well-defined viral sequences differing in their AT content. Firstly, the lowest denaturation temperature (T(p)) that allowed amplification of the parental sequence was determined. Secondly, differential amplification of AT-rich viral variants was obtained by using a denaturation temperature 1-3 degrees C lower than T(p). Application of this sensitive method to two different viruses allowed us to identify human immunodeficiency virus type 1 G-->A hypermutants in a situation where none were expected and to amplify AT-rich variants selectively within a spectrum of poliovirus mutants.
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https://hal-pasteur.archives-ouvertes.fr/pasteur-00013738
Contributor : Mireille Gau <>
Submitted on : Thursday, November 10, 2005 - 1:34:52 PM
Last modification on : Monday, January 13, 2020 - 5:08:10 PM

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Rodolphe Suspene, Michel Henry, Sophie Guillot, Simon Wain-Hobson, Jean-Pierre Vartanian. Recovery of APOBEC3-edited human immunodeficiency virus G->A hypermutants by differential DNA denaturation PCR.. Journal of General Virology, Microbiology Society, 2005, 86 (Pt 1), pp.125-9. ⟨10.1099/vir.0.80426-0⟩. ⟨pasteur-00013738⟩

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