Hepatitis C virus (HCV) infection can cause chronic liver disease that develops into cirrhosis and liver cancer. It is estimated that are more than 170 million of th world’s population suffering from HCV. Accurate diagnosis is needed to provide appropriate early treatmen, including preventing further transmission of the virus. The purpose of this study was to construct plasmid expression of recombinant antigen for detection of anti-HCV antibodies. The antigen coding gene is designed so that is composed to epitopes that are immunodominant, sustainable and and represent HCV subtypes circulating in Indonesia and globally. Furthermore, the gene was made by synthetic DNA techniques by DNA synthesis service provide... More
Hepatitis C virus (HCV) infection can cause chronic liver disease that develops into cirrhosis and liver cancer. It is estimated that are more than 170 million of th world’s population suffering from HCV. Accurate diagnosis is needed to provide appropriate early treatmen, including preventing further transmission of the virus. The purpose of this study was to construct plasmid expression of recombinant antigen for detection of anti-HCV antibodies. The antigen coding gene is designed so that is composed to epitopes that are immunodominant, sustainable and and represent HCV subtypes circulating in Indonesia and globally. Furthermore, the gene was made by synthetic DNA techniques by DNA synthesis service providers and accepted by the researchers in the form of blinding on the PUC57 plasmid to pQE80L plasmid with BamHI and HindIII cloning sites. Subcloned recombinant plasmids were then propagated on Top10 Escherichia coli cells and verified by PCR colony tecnique, restriction, and sequencing analysis. HCV recombinant antigen coding gene is 1200 bp. Cloning of these gene on the PUC57 vector produced a plasmid pUC57-HCV_ME (3910 bp) and subcloned in the pQE80L vector producing pQE80L-HCV_ME plasmid (5909bp). Based on verification results of pQE80L-HCV_ME plasmid the expression of recombinant antigen for detection of anti-HCV antibodies has been successfully constructed.
