Zell- und Molekularbiologie

Abstrakt

Molecular Study of Human HCV RNA by Real Time-Polymerase Chain Reaction, Viral Kit and Robogene Quantification

Taha Nazir

Hepatitis C virus (HCV) is one of the major causes of liver cirrhosis. It is highly mutable, circulates heterogeneously and potentially connected with diverse clinical phenotypes. Thus, we aim this study and collected blood samples from 183 HCV suspicious patients of age 31-58 years during March, 2013 to December, 2014. The prepared specimens were investigated for HCV RNA quantification by RT-PCR Robogene; isolation technique “Internal Virus Kit” (AJ Roboscreen, Germany). The subsequent samples were drawn after 4, 12 and 24 weeks. The reference range(s) of <15 IU/mL and <1.18 Log IU/mL were used to monitor therapy and/or disease progression. Levene’s test for Equality of Variances, t-test for Equality of Means and Fisher’s Exact tests were used for categorical variables to study the variance and measured association among variables. The specimens were collected from 183 HCV suspicious patients, age 31-58 years (Mean 44.83, SD 8.062). After amplification of the extracted RNA by RT- PCR using Bio-Rad’s CFX 96 Machine, 68 patients (37.15%) were identified/ diagnosed as HCV positive, 108 (59.02%) found negative and 7 (3.83%) patients did not responded to therapy of peginterferon and ribavirin. Paired samples statistical analysis showed significant variance in the mean values of HCV RNA quantification (IU/ ml) before therapy and at wk 12 (p value 0.055); before therapy and at wk 24 (P value 0.053). however, there is no significant variance seen Before therapy and at wk 4 (P value 0.082); at wk 4 and wk 12 (P value 0.106); at wk 4 and wk 24 (P value 0.101) and at wk 12 and wk 24 (P value 0.118). The quantification of human HCV RNA by RT-PCR, internal viral kit and Robogene quantification can potential be used to to rationalize the treatment, enhance antiviral responses and mitigate mortalities because of HCV RNA.