Optimization and application of subtype specific polymerase chain reaction for detection and identification of mixed subtypes of hepatitis C virus in patients with renal disease

Abstract

The main objective of the study is to optimize a subtype specific polymerase chain reaction (PCR) for identification of hepatitis C virus (HCV) subtypes in patients with renal disease. Thirty two peripheral blood specimens obtained from 28 patients (post renal transplant N=14, chronic kidney disease N=14) were subjected to HCV viral load determination followed by genotyping analysis. Based on the mixed genotypes and subtypes (>one subtype) obtained by type specific PCR, specific patterns from Pattern I through Pattern X were assigned. All the 32 peripheral blood specimens revealed mixed HCV subtype patterns (>one subtype). The detection of Pattern I in 12 (42.8%) out of 28 patients was statistically significant (Chi square test, P value<0.001). HCV subtyping assay developed using stringent thermal profile revealed the presence of mixed subtype patterns (>one subtype) which is for the first time being reported in literature.