title: Standardisation of primers and an algorithm for HIV-1 diagnostic PCR evaluated in patients harbouring strains of diverse geographical origin. The Belgian AIDS Reference Laboratories.
abstract: Eight Belgian AIDS Reference Laboratories established a multicentre quality control to evaluate the performance of their diagnostic human immunodeficiency virus type 1 (HIV-1) DNA polymerase chain reaction (PCR). A set of Belgian and African HIV-1 seropositive and seronegative patient samples, collected in Belgium, and the British Medical Research Council (MRC) HIV-1 PCR reference reagent kit, containing plasmid HIV-1 DNA at several dilutions in human carrier DNA with appropriate negative controls, were tested by the laboratories. No false positive results were reported. All laboratories were able to detect one to two copies of HIV-1 DNA. Among the 17 Belgian and African HIV-1 seropositives, some laboratories reported up to four indeterminate results, mainly due to failure of the SK38-39, SK68-69 (Ou et al. (1988) Science 239, 295-297) and/or gag881-882 (Simmonds et al. (1990) J. Virol. 64, 864-872) primers and a poorly performing algorithm. Only the H1POL4235-4538 nested pol primer set, developed by one of the laboratories, correctly identified all the tested HIV-1 positive and negative samples. Consequently, the laboratories decided to evaluate these pol primers as a reference primer set and to standardise the testing algorithm. All laboratories achieved a sensitivity and specificity of 100% on testing 10 additional Belgian and African patient samples, when adapting a standardised algorithm based on three HIV-1 primer sets, one of which is the H1POL4235-4538 primer set.
authors.affiliation: Katholieke Universiteit Leuven, Rega Instituut, Belgium.
authors.forename: A M
authors.initials: AM
authors.lastname: Vandamme
authors.forename: K
authors.initials: K
authors.lastname: Fransen
authors.forename: L
authors.initials: L
authors.lastname: Debaisieux
authors.forename: D
authors.initials: D
authors.lastname: Marissens
authors.forename: S
authors.initials: S
authors.lastname: Sprecher
authors.forename: D
authors.initials: D
authors.lastname: Vaira
authors.forename: A T
authors.initials: AT
authors.lastname: Vandenbroucke
authors.forename: C
authors.initials: C
authors.lastname: Verhofstede
mesh_terms: D000349:Africa; D000465:Algorithms; D001483:Base Sequence; D001530:Belgium; D017931:DNA Primers / Q000592:standards*; D004279:DNA, Viral / Q000097:blood; D005069:Evaluation Studies as Topic; D015751:Genes, pol / Q000235:genetics; D015658:HIV Infections / Q000175:diagnosis*; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000592:standards*; D011786:Quality Control; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D004279:DNA, Viral