AEGiS-15IAC: Genotypic analysis of the drug-resistance related point mutations in the pol-gene of HIV-I.

15th International AIDS Conference

Bangkok, Thailand - July 11-16, 2004

Genotypic analysis of the drug-resistance related point mutations in the pol-gene of HIV-I.

Int Conf AIDS 2004 Jul 11-16; 15:(abstract no. A10085)

Arora SK, Sachdeva N, Sehgal S, Datta U
PGIMER, Chandigarh, India

BACKGROUND: There are estimated 42 million cases of HIV worldwide as per WHO and with India contributing approximately 4.52 million cases. The infection is going un-abated in under-developed world due to limited access of the drugs to the infected population. With the gradual decrease in the cost of the anti-retroviral therapy (ART) comprising nucleoside and non-nucleoside reverse transcriptase inhibitors along with protease inhibitors. HIV genome replication being a highly error prone phenomenon, the emergence and transmission of drug-resistant mutants of HIV in the patients is known to be a major cause of treatment failure. Resistance to lamivudine (3-TC) occurs mainly due to point mutations at codon 184 (Met[right arrow]Val or Ile) in the RT region of HIV whereas resistance to Zidovudine (AZT) mainly involves codons 70 (Lys[right arrow] Arg) & 215 (Thr[right arrow]Phe). Therefore, it is important that such mutations be screened in order to determine the successful treatment strategies.

METHODS: The pol-gene fragment (768 bp) was first amplified from proviral DNA and mutations were detected by ARMS-PCR using wild and mutant codon-specific primers. The mutations in the pol-gene leading to 3-TC related resistance were screened in 60 treatment naïve & 10 treated patients.

RESULTS: The mutant variants were observed in 20 treatment naïve patients (Met Val in 4 and Met Val as well as Ile in 16 patients) while 2/10 treated patients also showed mutations. For AZT resistance the mutations at codon 215 & 70 was checked in 32 & 17 treatment naïve cases respectively. Codon 215 mutation could be detected in only one patient while mutation at codon 70 was more frequent and could be detected in 15/17 tested cases. CD4/CD8 cell counts by flowcytometry, Plasma HIV-RNA copy numbers by RT-PCR and soluble TNFRII levels were also determined in few cases. The correlation of the appearance of mutations vis-a-vis plasma viral load and CD4 counts is being further investigated.

Keywords: AEGIS, HIV-1, Genes, pol, Zidovudine, HIV-1 Reverse Transcriptase, Anti-HIV Agents, Point Mutation, Acquired Immunodeficiency Syndrome, Viral Load, HIV Infections, Drug Resistance, HIV, HIV Protease, Virus Replication, Codon, Mutation, HIV Seropositivity, Nucleosides, India, Humans, genetics, virology

040711
A10085