Quantification of Proviral Load of Human Immunodeficiency Virus Type 2 Subtypes A and B Using Real-Time PCR

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FONTE

American Society for Microbiology

RESUMO

We have developed and evaluated a new method to quantify human immunodeficiency virus type 2 (HIV-2) proviral DNA based on LightCycler real-time PCR. The assay has a detection limit of 5 copies/105 peripheral blood mononuclear cells (PBMC) and is insensitive to HIV-2 strain variability: HIV-2 subtypes A and B are both recognized and quantified. The intra- and interassay coefficients of variation range from 16 to 40% for high provirus concentrations (5 × 105 copies) and from 41 to 39% for low concentrations (5 copies). We used this method to compare the proviral DNA load and viral RNA load in plasma with clinical and immunological status for 29 patients infected by HIV-2 (subtype A in 17 and subtype B in 12). The proviral load (median, 201 copies/105 PBMC) was similar to that reported for HIV-1 infection. The median proviral loads did not correlate with the CD4+ cell count categories and were as follows for CD4+ cell counts of >400, 200 to 400, and <200 cells/mm3, respectively: 121 copies/105 PBMC (n = 8; range, <5 to 712 copies/105 PBMC); 114 copies/105 PBMC (n = 9; range, <5 to 1,907 copies/105 PBMC); and 285 copies/105 PBMC (n = 12; range, 53 to 2,524 copies/105 PBMC). Proviral load did not correlate with plasma HIV-2 RNA positivity. As HIV-2 is considered to replicate less efficiently than HIV-1, these high proviral loads might be explained by the proliferation of infected cells.

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