Stability in the HIV vDNA pool in peripheral CD4+ T cells of untreated patients by single tube quantitative PCR

• Published in: Journal of virological methods Vol. 87; no. 1; pp. 1 - 12
• Main Authors: Hockett, Richard D, Saag, Michael S, Kilby, J.Michael, Sfakianos, Greg, Wakefield, Theresa B, Bucy, R.Pat
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01-06-2000; Amsterdam Elsevier; New York, NY
• Subjects: AIDS/HIV; Biological and medical sciences; CD4+ T cells; CD4-Positive T-Lymphocytes - virology; disease progression; DNA; DNA, Viral - analysis; Follow-Up Studies; HIV Infections - blood; HIV Infections - virology; HIV vDNA; HIV-1 - isolation & purification; Human immunodeficiency virus; Human viral diseases; Humans; Infectious diseases; Longitudinal Studies; Medical sciences; Polymerase Chain Reaction - methods; Proviruses - genetics; Proviruses - isolation & purification; Reproducibility of Results; RNA, Viral - analysis; Single tube quantitative PCR; Viral diseases; Viral diseases of the lymphoid tissue and the blood. Aids; Viral Load - methods; HIV vDNA; Single tube quantitative PCR; CD4+ T cells; Human; Immunopathology; Stability; Retroviridae; AIDS; Immune deficiency; Lentivirus; Infection; Virus; Polymerase chain reaction; Investigation method; Viral disease; DNA; T-Lymphocyte; Human immunodeficiency virus
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(00)00139-7

HIV infection leads to loss of CD4 T cells and development of AIDS in most individuals without treatment. While disease progression during HIV infection correlates with the plasma viral load, much less is known about the levels of HIV vDNA. This paper describes the development and validation of a sensitive, quantitative PCR assay for the assessment of HIV vDNA. The system uses novel single tube, multiply competitive PCR technology, which allows five-point competitor competition in a single PCR reaction. The reproducibility and performance characteristics of the assay are extensively studied, which indicate that the system performs well in high DNA backgrounds. Using this assay system on a cohort of protease naı̈ve patients, HIV vDNA was assessed from PBMCs over an average follow-up period of 5 years. The data indicate that the HIV vDNA pool does not appreciably accumulate over the follow-up period, with many of the patients followed for up to 8 years. A reliable, quantitative assessment of vDNA pools will allow a better understanding of the dynamics of HIV pathogenesis.