Abstract
Infection with hepatitis C virus (HCV) is a major cause of chronic liver disease. On the other hand, Nosocomial transmission of HCV is a concern in haemodialysis (HD) units worldwide. In these patients, blood transfusions and long term dialysis are risk factors for transmission of HCV. Diagnosis of HCV infection is currently based on the detection of anti HCV antibodies by ELISA, and is confirmed by HCV RNA. The aim of the present study was to identify and detect the circulating non structural protein by using ELISA ,SDS-PAGE and Western blot techniques and to evaluate the usefulness of the detection of HCV antigen using ELISA for therapeutic follow-up (at 0 times, 12, 24 and 48 weeks) in selected 10 positive HCV-RNA patients at 0 times . Serum samples of 75 chronic hepatitis C (CHC) patients, serum samples of 75 haemodialysis patients and 25 healthy individual's sera as a negative control were included in this study. HCV antigen was detected in these samples using ELISA and Western blotting techniques. Western blot analysis showing a single immune reactive band in serum of CHC and haemodialysis patients infected with HCV at 27-KDa. ELISA technique was applied to detect the 27-KDa antigen. The cutoff level of ELISA above or below which the tested sera were considered positive or negative was calculated and was found to be 0.28 Based on this cut off the HCV antigen was detected in 80 % of CHC patients and in 36 % of haemodialysis patients. While, it was found that all healthy individuals used as a control were 100 % negative for HCV antigen. Furthermore, detection of HCV-RNA using nested PCR and HCV-NS4 antigen using ELISA for pre-therapeutic and therapeutic (combined interferon (IFN) and ribavirin therapy) at 0 time, 12, 24 and 48 weeks in 10 HCV-infected persons undergoing treatment was studied. The detection of HCV-NS4 antigen for these 10 positive HCV-RNA samples at (12 weeks and 24 weeks) showed that (60 %) were responsive for treatment and it was found that (40 %) did not respond.
Highlights
The global public health impact of chronic hepatitis C virus (HCV) infection and consequent liver disease continues to grow in numbers
The detection rate of HCV antigen was very obvious in chronic hepatitis C patients (80%) comparing with those of healthy individuals
The circulating non structural protein (NS4) was identified in sera from haemodialysis patients infected with HCV as well as chronic hepatitis C patients at 27 KDa which is similar to the HCV NS4B protein molecular weight (Konan et al, 2003)
Summary
The global public health impact of chronic HCV infection and consequent liver disease continues to grow in numbers. It has been estimated that there are over 170 million carriers of HCV worldwide with increasing incidence of new infections (Dennis et al, 2005). HCV is a major health problem in Egypt (Ray et al, 2000). It has the largest epidemic of hepatitis C virus in the world. A high prevalence of HCV infection in haemodialysis patients in addition to the risk factors such as the number of blood transfusions or duration on haemodialysis have been identified (Hinrichsen et al., 2002). The risk for HCV infection is dramatically increased during transfusion. Most of patients will progress to chronic hepatitis and some will even develop cirrhosis and hepatocellular carcinoma (Fung et al, 2009)
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