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5th International Workshop on HIV Drug Resistance & Treatment Strategies4-8 June 2001, Scottsdale, Arizona |
OBJECTIVE: Recent studies have demonstrated the potential for transferred resistant mutant virus in antiviral treatment-naïve patients. The work largely focused on blood compartments and relative mutational patterns of donor recipient pairs when available. Here, we describe viral genetic profiles of five patients in blood versus seminal compartments to identify mutant populations in a single patient that may be transferred during sexual contact.
METHODS: Genotypic resistance profiles from individual patients in two compartments, blood and seminal, were evaluated for specific resistance-associated mutations and polymorphic variation. Five patients with various treatment strategies were independently assessed for genotypic resistance in plasma-free virus, blood provirus, seminal plasma-free virus, and seminal cell associated provirus. Several extraction methods were used for targetted nucleic acids depending upon the source materials used (RNA or DNA). Evaluation of sequenced nucleotides was accomplished using the TRUGENE HIV-1 Genotyping kit and sequencing platform (Visible Genetics), which automatically assessed resistance-associated mutations and polymorphic variables as compared to a reference sequence.
RESULTS: Plasma-free virus, blood provirus and seminal plasma-free virus had similar resistance-associated mutations, which reflected past or currently failing drug regimens. Comparison of polymorphic variations not associated with drug resistance in these compartments (polymorphic fingerprint) was highly correlative. These results were consistent in all five patients tested. However, the seminal cell provirus, did not match genotypes obtained from the other genotyped compartments. One patient, who had been sequentially genotyped since 1994 in plasma and blood provirus compartments, had a seminal cell provirus in 2000, which matched the plasma genotype sequence from 1997. Because the cellular component of seminal fluid is genetically different in proviral content from other compartments tested, compartmentalized genotyping in seminal reservoirs relating to mutant transmission and salvage therapy should be examined.
PRESENTING AUTHOR: RM Lloyd Jr
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