MOLECULAR EPIDEMIOLOGY AND GENETIC CHARACTERIZATION OF HEPATITIS C VIRUS IN PATIENTS FROM SUBURBAN RAWALPINDI, P AKISTAN

Abstract

Hepatitis C virus is a devastating virus, known to mankind since 1989, but circulating in the human blood for centuries. It has affected millions of people from all ethnic origins, all walks of life and all socioeconomic statuses. Viral prevalence and incidence rates vary from country to country. Chronicity of the disease causes liver damage, functional impairment, liver shrinkage and hepatocellular carcinoma. Vinls disperses via blood and blood products. This study was basically conducted for identification of potential risk factors, their correlation with particular genotypes, their role in HCV transmission and evolutionary relationships of the genotypes. In order to accomplish the task 1200 patients from ethnic origins (residentials) of Gujar Khan, suburban Rawalpindi, Pakistan were randomly selected from two hospitals i.e. Pauji Foundation Hospital Rawalpindi and Pakistan Institute of Medical Sciences Islamabad, by convenient sampling method, for epidemiological studies. Futiher, plasma of 400 out of 1200 randomly selected studied participants was taken for RNA extraction. To find out the circulating genotypes in this population molecular reverse transcription PCR (RT peR), nested PCR and Restriction Fragment Length Polymorphism (RFLP) based analysis was performed. The genotypes were assigned to the samples by RFLP analysis and unresolved samples by RFLP were taken to sequencing level and 5' UTR was the target site for the sequencing. Sequenced samples were then cleaned, aligned and submitted to GenBank. Neighbour joining (NJ) method was used for phylogenetic analysis and rooted dendrograms were dragged to study the association between different viral types and subtypes. It was observed that majority of the studied participants, were chronic HCV carriers and married individuals are more likely to acquire the disease than unmarried individuals. There were high numbers of females; with major affected age group 40-49 years for both genders. The disease was related to the low socioeconomic status of the patients and low literacy levels in general; people from all profession were at equal risk of disease acquisition, with no additional risk for health care providers. Mostly reported risk factors in studied cohort were dental procedures, blood transfusions and therapeutic injections. There was no significant association between genotypes and any specific risk factor. In other words it will be more appropriate to say that all risk factors were equally contributing in the spread of HCV. Moreover, it was found that major