title: Low cost HIV-1 quantitative RT-PCR assay in resource-limited settings: improvement and implementation.
abstract: Monitoring of HIV viral load in low and middle income settings is limited by high cost of the commercial assays. Therefore, we developed a novel RT-PCR quantitative assay was developed. This assay targets the HIV-1 pol integrase gene (INT). Subsequently, the performance of the INT assay, described previously as a Long Terminal Repeat (LTR) assay and a combined INT/LTR dual target RT-PCR assay was compared. The LTR-assay was found to be sensitive and cost-effective (50-70% cheaper than commercial assays) with the lowest coefficient of variation (%CV). Introduction of an internal standard further improved assay reliability. Therefore, this LTR assay was implemented in West Java, Indonesia. Linearity and precision of the LTR assay were good: %CV ranged from 1.0% to 10.4%. The limit of quantitation was 616 copies/ml. Performance was comparable with the commercial assay (Abbott assay) (r(2)=0.01), although on average the viral loads were 0.39 log(10)copies/ml lower. In clinical practice, it had excellent capability for monitoring treatment failure, the positive predictive value was 99% and the negative predictive value was 93%. In conclusion, the implementation of the improved HIV-1 viral load LTR-assay for routine diagnosis in resource poor settings can be a good alternative when commercial assays are unaffordable.
authors: Department of Virology, Erasmus Medical Center, The Netherlands.
authors: Azzania
authors: A
authors: Fibriani
authors: Nadya
authors: N
authors: Farah
authors: Inri
authors: I
authors: Kusumadewi
authors: Suzan D
authors: SD
authors: Pas
authors: Reinout
authors: R
authors: van Crevel
authors: Andre
authors: A
authors: van der Ven
authors: Charles A B
authors: CA
authors: Boucher
authors: Martin
authors: M
authors: Schutten
mesh_terms: D003365:Costs and Cost Analysis; D016903:Drug Monitoring / Q000379:methods; D015658:HIV Infections / Q000175:diagnosis* / Q000821:virology; D019427:HIV Integrase / Q000235:genetics; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007214:Indonesia; D025202:Molecular Diagnostic Techniques / Q000191:economics / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000191:economics / Q000379:methods*
chemical_list: D019427:HIV Integrase