A multiplex real-time PCR for quantification of HIV-1 DNA and the human albumin gene in CD4+ cells

• Published in: APMIS : acta pathologica, microbiologica et immunologica Scandinavica Vol. 111; no. 6; pp. 625 - 633
• Main Authors: ERIKSSON, LARS E., LEITNER, THOMAS, WAHREN, BRITTA, BOSTRÖM, ANN-CHARLOTTE, FALK, KERSTIN I.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Munksgaard International Publishers 01.06.2003; Blackwell
• Subjects: Base Sequence; Biological and medical sciences; CD4-Positive T-Lymphocytes - immunology; CD4-Positive T-Lymphocytes - physiology; CD4-Positive T-Lymphocytes - virology; DNA load; DNA, Viral - genetics; Fundamental and applied biological sciences. Psychology; Genes, pol - genetics; HIV Infections - blood; HIV Infections - diagnosis; HIV Infections - virology; HIV-1; HIV-1 - genetics; HIV-1 - immunology; Human viral diseases; Humans; Infectious diseases; Longitudinal Studies; Medical sciences; Medicin och hälsovetenskap; Microbiology; Molecular Sequence Data; PI-ART; Polymerase Chain Reaction - methods; real-time PCR; Reproducibility of Results; RNA load; RNA, Viral - chemistry; RNA, Viral - genetics; Sequence Analysis, DNA; Serum Albumin - genetics; Techniques used in virology; therapy interruption; Viral diseases; Viral diseases of the lymphoid tissue and the blood. Aids; Viral Load; Virology; Human; Immunopathology; Multiplex polymerase chain reaction; HIV-1 virus; Retroviridae; Albumin; AIDS; Real time; Immune deficiency; Lentivirus; Infection; Virus; Treatment; Gene; Viral disease; DNA; Antiviral; Human immunodeficiency virus; Viral load; Quantitative analysis
• ISSN: 0903-4641; 1600-0463
• DOI: 10.1034/j.1600-0463.2003.1110605.x

We have established a simple system for measuring HIV‐1 DNA load in CD4+ cells. In a multiplex configuration, a conserved region in the HIV‐1 pol gene and a section of the human albumin gene were simultaneously amplified to estimate the number of HIV‐1 DNA copies per cellular genome. An established Epstein‐Barr virus (EBV) standard system was used to calibrate the HIV‐1 quantification. Our multiplex PCR system was tested on different in vitro developed HIV‐1 strains and on longitudinal samples from eight patients. The system was able to amplify both in vitro and in vivo samples of various genetic compositions. In all eight patients, HIV‐1 DNA was detected and ranged between 0.17 and 51×10−3 copies per CD4+ cell and could be monitored longitudinally, including long‐term PI‐ART and STI. The measured HIV‐1 DNA load may be used to select the best time for the institution or re‐institution of therapy.