STANDARDIZATION OF PRIMERS AND AN ALGORITHM FOR HIV-1 DIAGNOSTIC PCR EVALUATED IN PATIENTS HARBORING STRAINS OF DIVERSE GEOGRAPHICAL ORIGIN

Eight Belgian AIDS Reference Laboratories established a multicentre qu ality control to evaluate the performance of their diagnostic human im munodeficiency virus type 1 (HIV-1) DNA polymerase chain reaction (PCR ). A set of Belgian and African HIV-1 seropositive and seronegative pa tient 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 negat ive controls, were tested by the laboratories. No false positive resul ts were reported. Ah laboratories were able to detect one to two copie s of HIV-1 DNA. Among the 17 Belgian and African HIV-1 seropositives, some laboratories reported up to four indeterminate results, mainly du e to failure of the SK38-39, SK68-69 (Ou et al. (1988) Science 239, 29 5-297) and/or gag881-882 (Simmonds et al. (1990) J. Virol. 64, 864-872 ) primers and a poorly performing algorithm. Only the H1POL4235-4538 n ested pol primer set, developed by one of the laboratories, correctly identified all the tested HIV-1 positive and negative samples. Consequ ently, the laboratories decided to evaluate these pol primers as a ref erence primer set and to standardise the testing algorithm. All labora tories achieved a sensitivity and specificity of 100% on testing 10 ad ditional Belgian and African patient samples, when adapting a standard ised algorithm based on three HIV-1 primer sets, one of which is the H 1POL4235-4538 primer set.