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Performance evaluation of the two protease sequencing primers of the Trugene HIV-1 genotyping kit.

Ribas SG, Heyndrickx L, Ondoa P, Fransen K

Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium.

This article describes the performance of the two protease sequencing modules available in the Trugene HIV-1 genotyping Kit on a sample population with a high prevalence of HIV-1 non-B subtypes (n=110). The relevance of the algorithm recommended by the kit was also evaluated. The results indicated a high sequencing failure rate of the PR module (34%). Forty-five percent of the failed sequences derived from non-B subtype viruses. Furthermore, no PR sequence could be obtained from any of the HIV-1 subtype A and C infected samples that were tested. In contrast, a sequence could be obtained from the entire panel using the P2 module. The data indicated that the high rate of sequencing failures of the PR module was related to both the HIV-1 non-B subtypes as well as lower levels of RNA viral load. In six out of the 73 samples for which both protease modules were successful, discrepancies between the two protease sequences were observed, which led to discordant resistance reports in two cases. The data highlight the problems and the clinical implications that may occur during resistance genotyping of clinical samples with a high prevalence of HIV-1 non-B subtypes.

Published 20 June 2006 in J Virol Methods, 135(2): 137-42.
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