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  • Title: Evaluation of NucliSens-AmpliScreen methodology to detect subtypes G of HIV-1 and 4c/4d of HCV in the screening of blood donors.
    Author: Araújo F, Henriques I, Monteiro F, Meireles E, Cruz A, Tavares G, Mota-Miranda A.
    Journal: Transfus Clin Biol; 2005 Oct; 12(4):331-5. PubMed ID: 16182583.
    Abstract:
    BACKGROUND AND OBJECTIVES: Albeit, the NucliSens Extractor combined with the Ampliscreen was validated for application in NAT minipool screening, a study to evaluate the reliability of the procedure in relation to subtypes G of human immunodeficiency virus (HIV)-1 RNA and 4c/4d of hepatitis C virus (HCV) RNA should be performed, due to their genetic differences and the high frequency in our country. STUDY DESIGN: Samples from patients infected with subtypes G of HIV-1 RNA and 4c/4d of HCV RNA were diluted with negative plasma and tested eight times for each concentration. For nucleic acid extraction we used an automated silica-based extraction method (NucliSens Extractor) and for amplification and detection the AmpliScreen HIV-1 version 1.5 and AmpliScreen HCV version 2.0 (Roche Diagnostic Systems) were applied. RESULTS: The sensitivity for HIV-1 RNA genotype G using the NucliSens-AmpliScreen method-95% detection limit (95% CI) of 25 (18-50) copies per ml-is comparable with those described for genotypes B and E and to that obtained by the Multiprep procedure. In the case of HCV, the sensitivity of the method was also similar, when we compared the detection limits obtained for genotype 4c/4d-95% detection limit (95% CI) of 34 (24-71) IU/ml-with the genotype 1 published. CONCLUSIONS: The data presented here suggest that these infections will not be missed because of genetic variation, as the platform exhibited similar limits of detection for the subtypes evaluated, meeting the sensitivity requirements set by the regulatory bodies.
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