Screening of HIV-1 isolates by reverse heteroduplex mobility assay and identification of non-B subtypes in Italy

J Acquir Immune Defic Syndr. 2004 Oct 1;37(2):1295-306. doi: 10.1097/01.qai.0000123273.76723.04.

Abstract

Objective: The increasing prevalence of HIV-1 transmission through heterosexual contacts and the growing number of immigrants from non-Western countries, where non-B subtypes and recombinant forms are prevalent, suggest the possible emergence in Italy of a new epidemic wave of HIV-1 non-B subtypes as well as recombinant forms.

Methods: The distribution of HIV-1 subtypes has been evaluated in 63 seropositive individuals residing in Italy, most of whom were infected through a sexual route during the last 5 years. A modified heteroduplex mobility assay (HMA) strategy, reverse HMA (rHMA), has been developed in our laboratory, allowing rapid identification of divergent-from-B-subtype isolates, which have been subsequently characterized by detailed molecular and phylogenetic analyses.

Results: Five samples show, on rHMA, an electrophoretic pattern compatible with a non-B subtype classification. Their phylogenetic analysis, performed on both env and gag regions, confirms the rHMA subtyping prediction, given that 3 samples fall into the "A-family" subtype and 2 into the G subtype. The 5 non-B-subtype HIV-1 isolates have been identified among 23 variants (prevalence, 21.74%) isolated during the 2000 to 2001 period in heterosexuals. In parallel, B-subtype isolates show high levels of intrasubtype nucleotide divergence, compatible with a constant HIV-1 molecular diversification.

Conclusion: The Italian HIV-1 epidemic is still mostly attributable to the B subtype, which shows an increasing nucleotide heterogeneity. Heterosexual transmission and the interracial blending, however, are slowly introducing novel HIV-1 subtypes, and the data indicate that rHMA represents a powerful tool for HIV-1 biomolecular screening in epidemics characterized by a mono-/dual-subtype predominance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • DNA, Viral / analysis
  • DNA, Viral / blood
  • Female
  • Genes, env
  • HIV Infections / transmission*
  • HIV Infections / virology
  • HIV Seropositivity
  • HIV-1 / classification*
  • HIV-1 / genetics
  • HIV-1 / isolation & purification
  • Heteroduplex Analysis / methods*
  • Humans
  • Italy
  • Male
  • Nucleic Acid Heteroduplexes
  • Phylogeny
  • Sequence Analysis, DNA

Substances

  • DNA, Viral
  • Nucleic Acid Heteroduplexes