Prevalence of Hepatitis C Virus (HCV) Genotype 3a in the Infected Population of Lahore, Pakistan

Hepatitis C virus (HCV) genotype 3 is widely distributed, and genotype 3-infected patients achieve a lower cure rate in direct-acting antiviral (DAA) therapy and are associated with a higher risk of hepatic steatosis than patients with other genotypes. Thus, the study of the virology and pathogenesis of genotype 3 HCV is increasingly relevant. Here, we developed a full-length infectious clone and a subgenomic replicon for the genotype 3a isolate, CH3a. From an infected serum, we constructed a full-length CH3a clone, however, it was nonviable in Huh7.5.1 cells. Next, we systematically adapted several intergenotypic recombinants containing Core-NS2 and 5′UTR-NS5A from CH3a, and other sequences from a replication-competent genotype 2 a clone JFH1. Adaptive mutations were identified, of which several combinations facilitated the replication of CH3a-JFH1 recombinants; however, they failed to adapt to the full-length CH3a and the recombinants containing CH3a NS5B. Thus, we attempted to separately adapt CH3a NS5B-3′UTR by constructing an intragenotypic recombinant using 5′UTR-NS5A from an infectious genotype 3a clone, DBN3acc, from which L3004P/M in NS5B and a deletion of 11 nucleotides (Δ11nt) downstream of the polyU/UC tract of the 3′UTR were identified and demonstrated to efficiently improve virus production. Finally, we combined functional 5′UTR-NS5A and NS5B-3′UTR sequences that carried the selected mutations to generate full-length CH3a with 26 or 27 substitutions (CH3acc), and both revealed efficient replication and virus spread in transfected and infected cells, releasing HCV of 104.2 f.f.u. ml−1. CH3acc was inhibited by DAAs targeting NS3/4A, NS5A and NS5B in a dose-dependent manner. The selected mutations permitted the development of subgenomic replicon CH3a-SGRep, by which L3004P, L3004M and Δ11nt were proven, together with a single-cycle virus production assay, to facilitate virus assembly, release, and RNA replication. CH3acc clones and CH3a-SGRep replicon provide new tools for the study of HCV genotype 3.

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Prevalence of Genotype 3a in Different Regions of Pakistan: A systematic Review and Meta-Analysis

Journal of Pharmaceutical Research International ◽

10.9734/jpri/2021/v33i44a32628 ◽

2021 ◽

pp. 376-382

Author(s):

Rehan Ahmad Khan Sherwani ◽

Maria Aslam ◽

Kanwal Saleem ◽

Atif Khan Jadoon ◽

Hafiz Muhammad Nawaz

Keyword(s):

Systematic Review ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Literature Review ◽

Confidence Interval ◽

Meta Analysis ◽

Hcv Genotype ◽

Genotype 3A

The present study aims to explore the prevalence of genotype 3a under hepatitis C virus (HCV) infections among all the provinces of Pakistan. It is alarming to note that Pakistan stands in the second position for having a large number of cases of HCV every year. Six major genotypes characterize HCV. To study the overall prevalence of HCV and its associated genotype 3a in all the provinces of Pakistan, a systematic review and meta-analysis were performed using STATA version 14.2. The published studies conducted in all the regions of Pakistan reported the incidence of HCV genotype 3a were shortlisted. The pooled summary estimates were calculated along with their confidence interval by using the "Metaprop" command. The literature review showed that the prevalence of HCV genotype 3a is most common in all the provinces of Pakistan. It is revealed that the prevalence of HCV genotype 3a was 86.46% in Punjab, which is the highest among all the regions.

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Anti-viral effect of Indigoferra gerardiana on core protein of HCV and clinical significance of serum markers in chronic HCV genotype 3a patients in district Peshawar

10.21203/rs.2.23383/v1 ◽

2020 ◽

Author(s):

Shahina Mumtaz ◽

Jawad Ahmed ◽

Shafiq Ahmad Tariq ◽

Waheed Iqbal ◽

Sami Siraj ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Antiviral Activity ◽

Methanolic Extract ◽

Core Protein ◽

Categorical Variables ◽

Cross Sectional ◽

Hcv Genotype ◽

Wide Range ◽

Genotype 3A

Abstract Background/aim: Hepatitis C virus (HCV) disease is a health challenge due resistance cases globally. Many plant-derived natural compounds have demonstrated antiviral effects. These plants can provide an alternative way to new antiviral. Core protein of HCV has ability to interact with a wide range of viral and cellular proteins, including proto-oncogenes. Materials and Methods: In this cross-sectional study, total of 421 Hepatitis C Virus (HCV) positive patients were subjected by Polymerase Chain Reaction (PCR). The correlation of genotype with continuous and categorical variables was analyzed. Furthermore, antiviral activity of indigoferra gerardiana (IG) was tested against core protein of HCV. The amplified complete core protein of HCV genotype 3a was sequenced and cloned. IG was extracted and screened against core protein of HCV 3a genotype in cell line. Results: Genotype 3a was the common genotype type of HCV (n=363, 86%). Significant differences were observed in viral load among 3a genotype of HCV (< P 0.05). Frequently detected age group was 21-40 (59.4%) Sex and age were statistically associated with HCV infection (49% males and 37% in females). The results of RT-PCR demonstrated that methanolic extract of IG showed 57% reduction of infecting cells. Conclusions: Methanolic extract of IG showed good antiviral activity i.e. 57% reduction of infecting cells.

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Evaluation of triple antiviral therapy response in patients with Hepatitis C Virus (HCV) in interior Sindh, Pakistan (New antiviral therapy response in patients with HCV genotype 3a)

10.5176/2251-2489_biotech16.21 ◽

2016 ◽

Author(s):

Shameem Bhatti ◽

◽

Sobia Manzoor

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Antiviral Therapy ◽

Therapy Response ◽

Hcv Genotype ◽

Genotype 3A

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Hepatitis C virus genotype and its correlation with viral load in patients from Kathmandu, Nepal

The Journal of Infection in Developing Countries ◽

10.3855/jidc.10391 ◽

2020 ◽

Vol 14 (12) ◽

pp. 1470-1474

Author(s):

Bhavesh Kumar Mishra ◽

Uday Narayan Yadav ◽

Saroj Khatiwada ◽

Man Kumar Tamang ◽

Shivir Dahal ◽

...

Keyword(s):

Hepatitis C Virus ◽

Viral Load ◽

Hepatitis C ◽

High Viral Load ◽

Hcv Rna ◽

Virus Genotype ◽

Median Viral Load ◽

Hcv Genotype ◽

Hcv Genotypes ◽

Genotype 3A

Introduction: Knowledge about the distribution of hepatitis C virus (HCV) genotype and its correlation with viral load are important for the decision of treatment and the prediction of disease progression, however such information is very limited in Nepal. Here, we investigated the distribution of HCV genotypes and viral load for HCV-infected patients from Kathmandu, Nepal. Methodology: Ninety-six patients with HCV infection and not on antiviral therapy were enrolled from three different medical centers in Kathmandu valley, Nepal. Demographics were recorded and blood samples were collected. Plasma was separated and HCV RNA was extracted. Reverse transcriptase PCR (RT-PCR) was performed to measure the viral load, and virus genotype was determined. Results: Genotype 3a (n = 53, 55.2%) was the most prevalent, followed by 1b (n = 19, 19.8%), 1a (n = 18, 18.8%), 5a (n = 3, 3.1%), and mix types (n = 3, 3.1%). The median viral load for HCV genotype 1a was 770,942 IU/mL (IQR, 215,268-3,720,075), 1b was 700,000 IU/mL (IQR, 431,560-919,000), 3a was 1,060,000 IU/mL (IQR, 641,050-6,063,500), 5a was 673,400 IU/mL, and mixed was 6,428,000 IU/mL. A correlation between genotype and viral load was observed (p = 0.02), of which genotype 3a showed a high viral load. Conclusions: HCV genotypes 1a, 1b, 3a, and 5a were identified in Kathmandu, Nepal, and mixed genotype patients were observed in the patients studied. HCV genotype showed a correlation with viral load in patient plasma. This finding may contribute to the treatment and prevention of hepatitis C in Kathmandu, Nepal.

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Hepatitis C virus genotype 3a: Predictive factors associated with treatment response in patients of Peshawar.

The Professional Medical Journal ◽

10.29309/tpmj/2019.26.08.3876 ◽

2019 ◽

Vol 26 (08) ◽

pp. 1315-1322

Author(s):

Amina Gul ◽

Naheed Gul ◽

Ijaz Ali ◽

Jawad Ahmed

Keyword(s):

Hepatitis C Virus ◽

Viral Load ◽

Hepatitis C ◽

Treatment Response ◽

Virological Response ◽

Baseline Viral Load ◽

P Value ◽

Hcv Genotype ◽

Chronic Hcv ◽

Genotype 3A

Hepatitis C Virus (HCV) is a flavivirus responsible for causing chronic liver diseases including cirrhosis and hepatocellular carcinoma. Identification of various patient and virus-related factors that can help predict response to antiviral therapy is extremely important in formulating the best therapeutic strategy for each patient either to continue or stop the therapy. The present study aimed to determine Sustained Virological Response (SVR) in HCV genotype 3a infected patients that received combination therapy of Sofosbuvir and Ribavirin and to investigate various factors that can help predict SVR. Study Design: Longitudinal Study Settings: Institute of basic Medical Sciences, Khyber Medical University, Peshawar (IBMS, KMU). Period: July 2016 to September 2017. Materials and Methods: Treatment response was evaluated among 100 HCV genotype 3a infected patients that received Sofosbuvir and Ribavirin therapy for 24 weeks. Various baseline parameters including hematological, biochemical and viral profiles of were recorded. HCV genotype determination was carried out by type specific nested PCR based genotyping assay. Viral load was determined at baseline, at 12 weeks for Early Virological Response (EVR) and at 24 weeks of treatment for SVR. Viral RNA quantification was carried out by Real Time PCR. Results: Out of 100 patients, SVR was observed in 83% of patients; while 17% of the chronic HCV 3a infected patients were Non-Responders (NR). Mean age of patients was low 34 ± 9.8 among patients who achieved SVR as compared to patients with non-response (41 ± 10). The 24 weeks Alanine aminotransferase (ALT) levels were significantly lower among patients with SVR (p-value ≤ 0.05). Although statistically not significant, baseline viral load was higher in NR group (p-value ≥ 0.05), than those with SVR. Association of EVR with SVR was found statistically significant (OR= 2.8, 95% CI 1.2-6.4, p-value ≤ 0.05). Conclusions: The current study indicated that pre and on-treatment monitoring of patients receiving anti-viral therapy is important for the management of patients with chronic HCV infection.

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In VitroActivity of Daclatasvir on Hepatitis C Virus Genotype 3 NS5A

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01874-12 ◽

2012 ◽

Vol 57 (1) ◽

pp. 611-613 ◽

Cited By ~ 36

Author(s):

Chunfu Wang ◽

Lourdes Valera ◽

Lingling Jia ◽

Melissa J. Kirk ◽

Min Gao ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Chronic Infection ◽

Combination Therapies ◽

Genotype 3 ◽

Virus Genotype ◽

Replication Complex ◽

Hcv Genotype ◽

Potential Use ◽

Genotype 3A

ABSTRACTThe NS5A replication complex inhibitor daclatasvir (DCV; BMS-790052) inhibits hybrid replicons containing hepatitis C virus (HCV) genotype 3a (HCV3a) NS5A genes with 50% effective concentrations (EC50s) ranging from 120 to 870 pM. Selection studies with a hybrid HCV3a replicon identified NS5A residues 31 and 93 as sites for DCV-selected resistance. Our results support the potential use of DCV as a component in combination therapies for HCV3a chronic infection.

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