The Establishment Of Detecting The Genotyping Of Hepatitis C Virus Using DNA Chip With Coloration Methods

Background and objective: Hepatitis C virus(HCV), a member of the Flaviviridae family is the major cause of chronic liver disease worldwide. One of the important characteristics of HCV is that its genome exhibits significant genetic heterogeneity as a result of the accumulation of mutations during viral replication. The genetic sequences of HCV variants are very heterogeneous, varying by more than 30% across the entire genome among the six major genotypes, 20% among subtype, and up to 10% with a subtype. In this research, our objective is to develop a new DNA chip with coloration, which can be used for rapid and economical detection of the genotyping of hepatitis C virus(HCV).Methods: Seven plasmids, which contain seven kind viruses' specific DNA fragments, were constructed.Probes and primers were designed according to the sequence of HCV 5' non-coding region (5'NCR) to fabricate DNA chip. Experimental group consist of 60 positive serum samples and control group consist of 20 negative serum samples. To obtain the aimed gene, then they were hybridized with DNA chip. Finally, the result will be shown in a nylon film. The results of DNA sequencing of samples were used as the control in double blind experimental.Result: Detected using DNA chip, the serum specimens in the experimental group were all positive and the control group were all negative. The double blind test results showed a coincidence rate of 96.7% in genotyping of HCV by DNA chip and sequencing.Conclusion: It showed higher specialty and sensitivity using DNAchip to detect thegenotype of HCV;This technology was successfully applied to the genotyping of HCV.