Hepatitis B and C Virus Infections among Blood Donors in Blood Transfusion Center , Pokhara , Nepal : Seroprevalence and its Associated Risk Factors

Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) infections lead to chronic diseases and are the most common causes of liver cirrhosis and cancer in developing countries like Nepal. The study is carried out to determine the seroprevalence of HBV and HCV by using a Rapid kit method and Elisa Method to find out its risk factors. The cross-sectional study was done among blood donating people from 16th August 2016 to 19th November 2016. Blood donors in Pokhara Valley were screened for anti-HCV antibodies, anti-HBV antibodies using third generation ELISA kits and automated ELISA Processor in serology laboratory at Central Blood Transfusion Service (CBTS) of Nepal Red Cross Society (NRCS) in Pokhara, Nepal. 1777 (87.2%) units were male blood donors and 260 (12.6%) units were female donors out of 2037 participants. Gender wise, the ratio between male and female was 1:0.1. HBV and HCV infection rate in blood donors were detected at 0.7% (15/2037) and 0.5% (8/2037) respectively. HBV infection rate in volunteer blood donor people was 0.7% (14/1881) which was higher than the replacement donors i.e. 0.6% (1/156). Similarly, in HCV infection rate in volunteer donor were 0.4% (8/1881). HBV infected people are detected higher than the HCV infected people among the blood donors. In addition, there was no significant relationship between positive results of HBV and HCV tests with the gender, age, tattoo, donor type.


Introduction
Hepatitis B virus (HBV) is an enveloped, hepadnaviridae with partially double-stranded DNA [1].It is one of the cause of potentially lifethreatening liver infection and is a major global health problem.It can cause chronic liver disease and puts people at high risk of death from cirrhosis of the liver and liver cancer [2].Worldwide, an estimated two billion people have been infected with the Hepatitis B virus and more than 240 million have chronic (long-term) liver infections.In developing countries like Nepal, The highest prevalence of HBV was found in the 6-15 year age group [3].Common modes of transmission include perinatal, unsafe injection practices, unsafe blood transfusions, and unprotected sexual contact [4].Hepatitis C virus (HCV) is single-stranded RNA, enveloped flavivirus that causes liver disease [5].The disease can range in severity from a mild illness lasting for few weeks to a serious, lifethreatening condition that can lead from cirrhosis of the liver to liver cancer.About 170 million people are infected worldwide [6].HCV is most commonly transmitted through exposure to infectious blood and it may be transmitted through gender with an infected person or sharing of personal items contaminated with infectious blood, but these are less common [7].This study is conducted among blood donors in Pokhara Valley in order to study the HBV and HCV seroprevalence, among different groups and subgroups of blood donors.The main aim of this study is to determine the seroprevalence of HBV and HCV by Rapid kit and ELISA Method and find out its associated risk factors.Open land route facilitates the biological and virological spread of diseases [8].So, in the case of HCV and HBV, millions of migrant workers act as a biological transporter.However, documentation of HCV and HBV with past infection is frequent

Study Area and Population
Blood donors were selected according to the criteria of Blood Transfusion Service (BTS) based on the National Guidelines for BTS.The categories of donors that were present in this study were volunteers and replacement who may be the first time or repeated donor.Individuals selected by donor screening criteria as per NRCS' Standard Operating Procedure (SOP) of 2006 AD.
Participants were requested to fill a questionnaire form along with written consent were collected.Each donor was given an ID number for later trace out if needed.Inclusion criteria: Healthy donors were included for blood donation after taking an interview.The detailed inclusion criteria for blood donors are:donor should be in the age group between 18 to 60 years, body weight more than 45 kg for male and 35 kg for female, within normal range blood pressure 120/80 mm without medication and Hemoglobin should not be less than 12.6 g/ml.Exclusion criteria: Those who are found unhealthy on the interview were excluded for donation.The exclusion criteria for blood donors are:-in the case of a female, menstruating, pregnant and breastfeeding mother are not allowed for donation and Patients taking antibiotics.

Sample collection and processing
The blood bags were transported from blood donation campaign site to blood bank in igloo box containing ice bars to maintain the temperature near 4 0 C. A few drops of blood is used for blood grouping and 5ml of blood was allowed to clot or centrifuged at 3000 RPM for 2 minutes in a dry screw-capped test tube to separate serum samples to perform the test for HBV and HCV.

Detection of HBV and HCV by using Rapid kit test
Collected Sera were subjected for routine mandatory screening for HBsAg and anti-HCV antibodies by enzyme immunoassay-based rapid tests according to the standard protocol described by respective company (Hepacard, J. Mitra and Co., New Delhi, India, and HCV TRI-DOT, J. Mitra and Co., New Delhi, India) [10].

Statistical Analysis
Pearson's Chi-square test was used to determine the significant association of dependable variables.WinPepi software (version 11.65) was used for quantitative data analysis.A p-value < 0.05 was considered to be statistically significant at 95 % CI.

Results
A total of 2037 blood samples were collected from donors.The blood was screened by Rapid test and ELISA test to detect HBV and HCV infection.Gender wise, the ratio between male and female was at 1:0.1.The infection rate of HBV and HCV in blood donors were detected to be 0.7 % (15/2037) and 0.5% (8/2037) respectively (Figure 1).The infection rate in a male for HBV is 0.7% (13/1777) which was higher than HCV 0.4% (8/1777).Similarly, the infection rate in the female for HBV is 0.7% (2/260) which was also higher than HCV (0.0%) but no statistically significant, (p=0.280)result was obtained (Table 1).
HBV infection rate of married blood donors was 0.9% (12/1262) whereas 0.3% (3/775) was found in unmarried blood donors.Similarly, HCV infection rate of married blood donors was 0.7% (7/1262) whereas 0.1% (1/775) was found in unmarried blood donor with p=0.65 (Table 4).HBV infection rate was 0.7% higher in people without tattoo whereas the HCV infection rate in people without tattoo was 0.3% higher.Because in our case, the number of participants without tattoo was higher than the number of blood donors with tattoo.Although, it is widely known fact that cosmetic procedure, tattooing on a body can result in a HBV and HCV infection if the needles and equipment used are not properly sanitized.Besides, the poor tattooing procedure may have chances of microbial contamination.HBV and HCV infection rate in married blood donors were 0.9% and 0.7% respectively.Researchers concluded the marital status is a significant risk factor [18].They detected HBV and HCV were found 1.7% and 1.0% respectively in a married participant.HBV and HCV can be transferred from unprotected sexual contact if their partner is infected with the hepatitis virus.
People with unsafe sexual relationships and addiction to injectable drug users should always be careful as the chances of the virus getting transmitted are high among them.This severity of cases may be due to the lack of vaccination information in the population.The replication of HBV and HCV in HIV patients should be actively monitored while receiving antiviral therapy and this monitoring system should be made a part of clinical care [17].In the context of Nepal, people have has less knowledge of vaccination and those who knows avoid it because of hepatitis vaccination cost and no. of clinic visits to complete vaccination doses.The implications of screening tests for HBV or HCV co-infection in HIV patients are of great need in Nepal [18].

Limitations of the study:
This study was carried out in a limited population and only involved the donors attending on Mobile Health camp organized by BTS-NRS.So, the results obtained from this study might not be enough to indicate the actual burden of the disease in the large population.
Still, the study found the existence of endemic viral hepatitis among the people and need health policies to reduce the burden of viral hepatitis in society.PCR based detection method was not incorporated due to feasibility issues regarding time and resources constrain.

Conclusions
HBV and HCV infections can be transfer by piercing needles, general contact, blood

Figure 1 :
Figure 1: Infection of Hepatitis B Virus and Hepatitis C Virus in blood donors (Confirm by ELISA Method)

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