sensitive PCR assay system for the quantitation of viral genome equivalents: human immunodeficiency virus type 1 (HIV-1) and hepatitis B virus (HBV)

• Published in: Archives of virology 1997-01, Vol.142 (7), p.1297-1306
• Main Authors: Hämmerle, T, Himmelspach, M, Dorner, F, Falkner, F. G
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; disease course; DNA Primers; Electrophoresis, Polyacrylamide Gel; fluorescence; Fundamental and applied biological sciences. Psychology; gels; Genes, gag; genome; Genome, Viral; Hepatitis; Hepatitis B - virology; Hepatitis B virus; Hepatitis B virus - genetics; Hepatitis B virus - isolation & purification; HIV; HIV-1 - genetics; HIV-1 - isolation & purification; Human immunodeficiency virus; Human immunodeficiency virus 1; Humans; Microbiology; polyacrylamide; Polymerase chain reaction; Polymerase Chain Reaction - methods; quality control; Reference Standards; Reproducibility of Results; reverse transcriptase polymerase chain reaction; RNA, Viral - blood; Sensitivity and Specificity; Techniques used in virology; therapeutics; Viral Core Proteins - genetics; Viremia; Virology
• ISSN: 0304-8608; 1432-8798
• DOI: 10.1007/s007050050161

A sensitive and reliable quantitative method based on the polymerase chain reaction (PCR) and the reverse transcription polymerase chain reaction (RT-PCR) to detect and quantify human immunodeficiency virus (HIV-1) and hepatitis B virus (HBV), respectively, was developed. The samples are co-processed together with two internal standards (calibrators). The amplicons are separated on denaturing polyacrylamide gels and co-detected and quantitated by laser induced fluorescence. HIV-1 and HBV containing biological samples, including samples from international test panels, were accurately quantitated. The procedure has proven to be a valuable tool in the quality control of biologicals such as plasma products and may serve to monitor disease progression and response to antiviral therapy.