594-T. Performance of the TRUGENE HIV-1 Genotyping Kit and the Applied Biosystems HIV-1 Genotyping System in Sequence-Based Analysis of Non-B HIV-1 Subtypes

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Session 78 Poster Session
Diagnostic Techniques: Viral Sequencing Technologies
Session Time: 4:30-6:30 pm
Room 4E-F

594-T.

Performance of the TRUGENE HIV-1 Genotyping Kit and the Applied Biosystems HIV-1 Genotyping System in Sequence-Based Analysis of Non-B HIV-1 Subtypes
L. Jagodzinski*¹, J. Cooley¹, S. Kelly¹, and N. Michael²
¹Henry M. Jackson Fndn. and ²Walter Reed Army Inst. of Res., Rockville, MD

Background: Sequence-based genotypic assays that determine drug resistance mutations in the HIV genome are increasingly being used to make changes in patients’ drug regimens. The Visible Genetics’ TRUGENE HIV-1 Genotyping Kit is the first sequence-based assay to received FDA approval for use in patient management, making HIV-1 genotyping analysis a standard-of-care test in HIV-1 treatment decisions. Given the diversity of HIV-1 subtypes, the emergence of recombinant HIV-1 subtypes and the emergence of non-B subtypes in the United States, genotypic assays must be able to deliver sequence data on diverse HIV-1 subtypes.
Methods: A panel of 34 HIV-1 viral isolates representing HIV-1 subtypes A through H was used to assess the performance of 2 commercially manufactured assays: Visible Genetics’ TRUGENE HIV-1 Genotyping kit and Applied Biosystems’ HIV Genotyping System (ViroSeq).
Result: Visible Genetics’ TRUGENE assay successfully genotyped 94.1 % (32/34) of the HIV-1 subtype panel. Sequence data was not obtained for 1 C and 1 G subtype isolate in the TRUGENE assay. Applied Biosystems’ ViroSeq assay successfully genotyped 100% of the HIV-1 subtype panel. Comparison of nucleotide sequences generated by the 2 assays showed only minor differences that were related to single defined base vs mixed base calls. The inability of the TRUGENE assay to sequence 2 members of the HIV-1 subtype panel may indicate some problems in the amplification of the HIV-1 protease and reverse transcriptase gene regions. Clinical specimens when tested in both assays have failed in one or the other assay with a success rate of ~90% observed for both assays.
Conclusions: The Visible Genetics and Applied Biosystems’ HIV-1 Genotyping tests generated equivalent sequence data for all sequenced members of the HIV-1 subtype panel. Both tests are capable of delivering sequence data on diverse HIV-1 subtypes and detecting defined drug resistance mutations.