Descamps D, Masquelier B, Izopet J, Calvez V, Chaix ML, Ruffault A, Tamalet C, Yerly S, Brun-Vezinet F, Costagliola D; The ANRS Resistance Group; IAS Conference on HIV Pathogenesis and Treatment (2nd : 2003 : Paris, France).

Antivir Ther. 2003; 8 (Suppl.1): abstract no. 820.

CHU Bichat-Claude Bernard, Paris, France

BACKGROUND: To evaluate the performances of virology laboratories for HIV genotyping. METHODS: A coded panel of 10 samples was distributed to 38 laboratories and consisted of four diluted plasma samples from HIV-1-infected patients, five supernatants from cultivated viruses and one plasma HIV-negative control. Participants reported the amino acid at codons associated to NRTI, NNRTI and PI resistance listed on the web site: www.iasusa.org (2001). The reference amino acid sequences were defined as the most frequently reported by the participants. RESULTS: Thirty-eight laboratories reported datasets. All laboratories used automated sequencing technologies: Trugene (11), ViroSeq (2), ANRS consensus technique (20), in-house (5). Two laboratories amplified the negative control for both genes and one laboratory only the protease gene. All specimens were subtype B. The viral load of the nine positive specimens ranged between 283 and >750000 copies/ml. Two specimens with low viral load (283 and 1190 copies/ml) were not amplified for protease by three and five laboratories, and for RT by two and one, respectively. Seven samples had a high number of mutations in the protease (median 10, range 6-10) and in the RT (median 8, range 6-11). Two specimens had two and three mutations in the protease and no mutation in the RT genes. The global rate of false-positive and -negative results was 0.31% (36/11449) and 1.58% (70/4434), respectively. The frequency of false-positives was comparable in the RT (0.27%) and protease (0.38%) genes (P=0.3094), but the rate of false-negative was higher in the protease (1.95%) than in the RT (1.14%) (P=0.0390). The specimen in duplicate was scored identically by 89% (34/38) of the laboratories. CONCLUSION: The use of supernatants from cultivated viruses does not settle the question of determining which is the reference sequences as compared to the use of diluted plasma specimens. Quality control assessment for HIV drug resistance sequencing must include an HIV-negative control.

Publication Types:

• Meeting Abstracts

Keywords:

• Acquired Immunodeficiency Syndrome
• Drug Resistance
• Drug Resistance, Viral
• HIV Infections
• HIV Seropositivity
• HIV-1
• Humans
• Laboratories
• Mutation
• Quality Control
• Viral Load
• genetics
• methods
• reverse transcriptase, Human immunodeficiency virus 1

Other ID:

• GWAIDS0023475

UI: 102263099

From Meeting Abstracts