AAC
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by García Lerma, J. G.
Right arrow Articles by Heneine, W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by García Lerma, J. G.
Right arrow Articles by Heneine, W.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, February 1999, p. 264-270, Vol. 43, No. 2
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A Rapid Non-Culture-Based Assay for Clinical Monitoring of Phenotypic Resistance of Human Immunodeficiency Virus Type 1 to Lamivudine (3TC)

J. Gerardo García Lerma,1 Raymond F. Schinazi,2 Amy S. Juodawlkis,2 Vincent Soriano,3 Yulin Lin,2 Kathleen Tatti,2 David Rimland,2 Thomas M. Folks,1 and Walid Heneine1,*

HIV and Retrovirology Branch, Division of AIDS, STD, and TB Laboratory Research, Centers for Disease Control and Prevention, Atlanta, Georgia1; Georgia Veterans Affairs Research Center for AIDS and HIV Infections, and Department of Pediatrics and Department of Medicine, Emory University, Decatur, Georgia2; and Servicio de Enfermedades Infecciosas, Hospital Carlos III, Instituto de Salud Carlos III, Madrid, Spain3

Received 6 July 1998/Returned for modification 29 August 1998/Accepted 31 October 1998

Monitoring for lamivudine (3TC) resistance is important both for the clinical management of human immunodeficiency virus type 1 (HIV-1)-infected patients treated with 3TC and for surveillance of transmission of 3TC-resistant HIV-1. We developed a novel non-culture-based assay for the rapid analysis of phenotypic resistance to 3TC of HIV-1 in plasma. The assay measures the susceptibility of HIV-1 reverse transcriptase (RT) activity to 3TC triphosphate (3TC-TP) in plasma. RT detection was done by the Amp-RT assay, an ultrasensitive PCR-based RT assay. Under our assay conditions, we found that 5 µM 3TC-TP inhibited RT activity from wild-type (WT), zidovudine-resistant, or nevirapine-resistant HIV-1 but not from HIV-1 carrying either the M184V mutation or multidrug (MD) resistance mutations (77L/116Y/151M or 62V/75I/77L/116Y/151M). Mixing experiments showed a detection threshold of 10% 3TC-resistant virus (M184V) in a background of WT HIV-1. To validate the assay for the detection of phenotypic resistance of HIV-1 to 3TC in plasma samples, HIV-1 RT in 30 plasma specimens collected from 15 patients before and during therapy with 3TC was tested for evidence of phenotypic resistance by the Amp-RT assay. The results were compared with those of genotypic analysis. The RT in 12 samples was found to be 3TC sensitive, while the RT in 18 samples had evidence of phenotypic resistance. All 12 samples with 3TC-sensitive RT had WT genotypes at codon 184 and were retrieved before treatment with 3TC. In contrast, all 18 specimens with 3TC-resistant RT were posttherapy samples. This assay provides a simple, rapid, and reliable method for the detection of phenotypic resistance of HIV-1 to 3TC in plasma.

* Corresponding author. Mailing address: HIV and Retrovirology Branch, Centers for Disease Control and Prevention, 1600 Clifton Rd., NE, MS G-19, Atlanta, GA 30333. Phone: (404) 639-0218. Fax: (404) 639-1174. E-mail: WMH2{at}cdc.gov.

Antimicrobial Agents and Chemotherapy, February 1999, p. 264-270, Vol. 43, No. 2
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Clin. Vaccine Immunol. Clin. Microbiol. Rev.
J. Clin. Microbiol. ALL ASM JOURNALS

Copyright © 1999 by the American Society for Microbiology. All rights reserved.