title: Quantification of HIV-1 RNA in plasma: comparable results with the NASBA HIV-1 RNA QT and the AMPLICOR HIV monitor test.
abstract: We investigated and compared the reproducibility, accuracy, detection limits, and dynamic ranges of two commercial kits for quantification of RNA viral load using a titrated virus stock (laboratory strain HIV-1 IIIB) and 107 plasma samples of 25 HIV-1-infected patients. The high reproducibility of both methods (SD = 0.2-0.3 log for both methods) allowed reliable detection of a 0.5 log change in RNA viral load. Both methods had a similar detection limit (at least 10(3) RNA copies/ml plasma) and a dynamic range that extended over a 5 log (AMPLICOR) or a 6 log (NASBA) range of HIV-1 input. For HIV-1 IIIB, the viral load was compatible with measurements of virus-associated p24 antigen. For 21 patients (91 samples), the RNA viral load was similar with both methods differing by no more than 0.5 log. For four patients, the difference in viral load between the two methods was > 0.5 log for all 16 samples. For three of these patients, this could be explained by mismatches with primers or probes in the gag sequence: there was no correlation to the viral subtype. The RNA viral load determination was highly sensitive compared with p24 antigen measurement (> 95% of patients had a detectable viral load vs. 40% who had a detectable p24 level), but in the p24-positive samples the correlation between the antigen level and the RNA viral load was of only borderline significance. We also found that the viral RNA in whole blood was stable for at least 48 h during transport at room temperature. These observations show that both the NASBA HIV-1 RNA QT test and the AMPLICOR HIV monitor test are reliable parameters of the viral load, with great promise for their use as potential surrogate markers.
authors.affiliation: Rega Institute for Medical Research and University Hospitals, Katholieke Universiteit Leuven, Belgium.
authors.forename: A M
authors.initials: AM
authors.lastname: Vandamme
authors.forename: J C
authors.initials: JC
authors.lastname: Schmit
authors.forename: S
authors.initials: S
authors.lastname: Van Dooren
authors.forename: K
authors.initials: K
authors.lastname: Van Laethem
authors.forename: E
authors.initials: E
authors.lastname: Gobbers
authors.forename: W
authors.initials: W
authors.lastname: Kok
authors.forename: P
authors.initials: P
authors.lastname: Goubau
authors.forename: M
authors.initials: M
authors.lastname: Witvrouw
authors.forename: W
authors.initials: W
authors.lastname: Peetermans
authors.forename: E
authors.initials: E
authors.lastname: De Clercq
authors.forename: J
authors.initials: J
authors.lastname: Desmyter
mesh_terms: D001483:Base Sequence; D018450:Disease Progression; D005784:Gene Amplification; D016655:HIV Core Protein p24 / Q000032:analysis / Q000235:genetics / Q000276:immunology; D015658:HIV Infections / Q000097:blood / Q000209:etiology* / Q000276:immunology; D054303:HIV Reverse Transcriptase / Q000235:genetics; D015497:HIV-1 / Q000235:genetics* / Q000254:growth & development / Q000276:immunology / Q000472:pathogenicity; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000032:analysis* / Q000276:immunology; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load
chemical_list: D016655:HIV Core Protein p24; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; D054303:HIV Reverse Transcriptase