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Session 98 Poster Abstracts
Drug Resistance Testing: New Methods, Interpretations, and Reproducibility
Monday, 1:30 - 3:30 pm
Poster Hall


699
Performance of the Celera Diagnostics ViroSeq HIV-1 Genotyping System v2.0 for Sequence-based Analysis of Diverse HIV-1 Strains
S H Eshleman*1, J Hackett Jr2, S P Cunningham1, B Drews3, P Swanson2, C Brennan2, S G Devare2, L Zekeng4, L Kaptue5, and N Marlowe3
1Johns Hopkins Med. Inst., Baltimore, MD, USA; 2Abbott Labs., Abbott Park, IL, USA; 3Celera Diagnostics, Alameda, CA, USA; 4Lab. de Santé Hygiène Mobile, Yaoundé, Cameroon; and 5Univ. of Yaoundé, Cameroon

Background:  The Celera Diagnostics ViroSeq HIV-1 Genotyping System is an FDA-cleared, integrated system for sequence-based analysis of drug resistance mutations in subtype B HIV-1 protease and reverse transcriptase (RT). We evaluated the performance of this system for analysis of diverse HIV-1 strains.

Methods:  Plasma samples were obtained from 126 individuals from Uganda, Cameroon, South Africa, Argentina, Brazil, and Thailand with viral loads ranging from 2.92 to >6.0 log10 copies/mL. HIV-1 genotyping was performed using the ViroSeq system. HIV-1 subtyping was performed using phylogenetic methods.

Results:  PCR products suitable for sequencing were obtained for 125 (99%) of the 126 samples. Genotypes including protease (amino acids 1-99) and RT (amino acids 1-321) were obtained for 124 (98%) of the samples. Full bi-directional sequence data was obtained for 95 of those samples. The sequences were categorized into the following subtypes: A1/A2 (16), B (12), C (13), D (11), CRF01  AE (9), F/F2 (9), G (7), CRF02  AG (32), H (1), and intersubtype recombinant (14). Performance of the individual sequencing primers was examined. Genotyping of duplicate samples in a second laboratory was successful for 124 of the 126 samples. The homology of the sequence data from two laboratories ranged from 98% to 100% (median 99.8%).

Conclusions:  The ViroSeq system performs well for analysis of plasma samples with diverse non-B subtypes. Availability of this genotyping system should facilitate studies of HIV-1 drug resistance in non-subtype B strains of HIV-1.

Keywords: Drug resistance; subtype; genotyping