Detection of Human Immunodeficiency Virus Type 1 Clinical Isolates with Reduced Sensitivity to Zidovudine and Dideoxyinosine by RNA· RNA Hybridization

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 88; no. 8; pp. 3092 - 3096
• Main Authors: Japour, Anthony J., Chatis, Pamela A., Eigenrauch, Heidi A., Crumpacker, Clyde S.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 15.04.1991
• Subjects: AIDS; Allergic diseases; Amino acids; Antivirals; Biological and medical sciences; Cell lines; Clinical trials; General aspects; Genetic mutation; HIV; HIV 1; Immunopathology; Inhibitory concentration 50; Medical sciences; RNA; Human; Molecular hybridization; Immunopathology; RNA; HIV-1 virus; Retroviridae; AIDS; Hemopathy; Infection; Virus; Resistance; Autoradiography; Viral disease; Lentivirinae; Inosine; Human immunodeficiency virus; Quantitative analysis
• ISSN: 0027-8424; 1091-6490

A quantitative rapid assay to detect resistant clinical human immunodeficiency virus type 1 (HIV-1) strains remains an important medical goal. A system incorporating a quantitative RNA·RNA hybridization assay that measures the amount of intracellular HIV-1-specific RNA has been employed to detect the level of inhibition by nucleoside analogues in sensitive and resistant HIV-1 strains. The RNA·RNA hybridization assay readily distinguished previously published zidovudine (ZDV; 3'-azido-3'-deoxythymidine)-resistant isolates from ZDV-sensitive isolates of HIV-1. The 50% inhibitory concentration (IC50) of ZDV for HTLV-IIIBand sensitive clinical HIV-1 isolates is between 0.01 and 0.04 μM. HIV-1 strains from three patients on long-term ZDV therapy displayed a greater than 20-fold increase in the ZDV IC50compared to sensitive strains. The drug sensitivity system was confirmed by showing that mutations in the HIV reverse transcriptase gene from a ZDV-resistant isolate resulted in four amino acid changes (Leu-125 → Trp, Ile-142 → Val, Thr-215 → Tyr, and Pro-294 → Thr) including one change (Thr-215 → Tyr) that has been previously reported to be associated with resistance. One clinical HIV strain with high-level ZDV resistance displayed a 5-fold increase in 2',3'-dideoxyinosine IC50compared to that of HTLV-IIIB. A drug sensitivity assay employing RNA·RNA hybridization may be useful for extensive screening of HIV isolates from patients enrolled in clinical trials and permit the correlation of in vitro resistance with clinical outcome.