Validation of a nested PCR assay UMELOSA HCV CUALITATIVO for the detection of Hepatitis C virus.

An analysis of the performance characteristics of the UMELOSA HCV CUALITATIVO assay for the detection of Hepatitis C virus (HCV) RNA in human serum or plasma is presented. This qualitative in vitro diagnostic test is performed in three steps: (a) extraction of viral RNA, (b) reverse transcription of target RNA to generate complementary DNA followed by a polymerase chain reaction assay coupled with a second round of amplification, and finally (c) fluorescent detection of the amplified DNA by hybridization in a solid phase of an ultramicroplate coated with a complementary to amplified DNA probe. Considering the assay as a limit test for the control of impurities, the following analytical performance parameters were evaluated: specificity, detection limit and robustness.