In-Vitro Transcription analysis of NS5A from HCV-3a circulating in Pakistani patients with chronic hepatitis C and their differential response to antiviral therapy.

OBJECTIVE: Mutations in HCV nonstructural protein 5A (NS5A) play a vital role in virus resistance. The aim of this study was to develop a correlation between NS5A mutations (genotype 3a) and virological response towards interferon alpha (IFN-α) plus ribavirin therapy. METHODS: In this study, which was conducted from 09-02-2013 to 25-11-2015 in the rural area of Province Sindh - Pakistan, total patients' responses to peg-IFN therapy were investigated. Patients were given peg-IFN therapy for 24 to 48 weeks and categorized as sustained virologic responders (SVR) or non-responders (NR) to HCV infection. HCV NS5A region (2215-2335) of genotype 3a was identified in both responders and non-responders. RESULTS: Twenty-four NR with 24 SVR isolates showed significant mutations within the nonstructural protein 5A region in HCV genotype 3a. The New Zealand (NZL1) (GenBank D17763) differences were observed by using gene. The ISDR mutations for nonstructural protein 5A in non-responders have been reported as a possible explanation of HCV interferon resistance. CONCLUSION: Based on these results, it is suggested that decreased SVR is caused by the increased mutations in nonstructural protein 5A sequences. When the sequence outside the Protein kinases R binding domain (PKRBD) (2281-2335) was examined, significant differentiations were observed among the SVR and NR classes at few amino acid strains.

Molecular epidemiology and clinical features of Hepatitis C Virus (HCV) in epidemic areas of Interior Sindh, Pakistan.

OBJECTIVE: Highly variable genome of HCV and high prevalence in many geographical areas made it necessary to conduct local population studies. This study has been conducted to show HCV parameters along with clinical features in the local population of interior Sindh, province of Pakistan. METHODS: Present study was conducted in from August 2010 to November 2015 in the rural areas of Sindh, Pakistan. All the 31560 screened samples selected for the study were tested by second Generation Enzyme Linked Immunosorbent Assay (ELISA Biokit 480&96). RESULTS: Total 31560 people were screened for HCV and out of these 13.67% (n= 4314) HCV infected patients. When 4314 samples of patients were examined; the anti-HCV was significantly higher in males 2814 (14.98%) than in females 1500 (11.74%) with P value = 0.06. The age of the patients ranged from 18 to 65 years. Out of 4314 HCV samples, 3020 (70%) were of Genotype 3a, 237(5.5%) of Genotype 2a, 108 (2.5%) of Genotype- 1a, 216 (5%) of Genotype 1b, 237 (5.5%) of Genotype 3b and 43 (1%) of Genotype 4. Additionally, 108 (2.5%) had co-infection and 345 (8%) samples showed no result -designated as untypable by the genotyping. CONCLUSION: This study showed that HCV is most frequently reported disease with genotype 3a being the most prevalent genotype.

Correlation of OAS1 gene polymorphism at exon 7 splice accepter site with interferon-based therapy of HCV infection in Pakistan.

The most useful treatment for HCV infection worldwide is peg-interferon plus ribavirin, although the response varies from person to person. Hence, host genetics are significantly involved in the treatment response to HCV infection. The 2'-5' oligoadenylate synthetase (OAS) is one of the most important components of the immune system having significant antiviral functions. The aim of this study was to investigate the role of single nucleotide polymorphism (SNP) at the exon 7 splice acceptor site (SAS) of OAS1 to interferon-based therapy of HCV infection. OAS1 genotyping was performed in 140 HCV patients by restriction fragment length polymorphism polymerase chain reaction method (RFLP-PCR). These patients were enrolled for the study in 2010-2013. OAS1 SNP was also established in 120 healthy controls. Correlation of HCV genotypes, OAS1 SNP, and other factors with response to interferon therapy were statistically analyzed by SPSS 13 software. There were no significant differences in the distribution of OAS1 genotypes between healthy and patients subjects. The distribution of AG and AA genotypes of OAS1 genotypes between sustained virological responders (SVRs) and the non-responders (NRs) group were also comparable. However, Pearson chi square analysis indicated that the patients possessing a GG genotype of the OAS1 gene at exon 7 SAS demonstrated significantly positive association with treatment response to HCV infection (p=0.039). This study determined that SNP at exon 7 SAS of OAS1 was significantly associated with response to interferon-based therapy of HCV infection in our population.