Prepared Of Hepatitis Virus (HBV, HCV) Oligochip And Experimental Study Of A New Hepatitis Virus SENV

HBV and HCV are the most common and harmful hepatitis viruses, the chronic infection of which are closely related to the development of cirrhosis and hepatocellular carcinoma (HCC). In our country, HBV is high prevalence, the carriers of HBV are more than one hundred million at least, more than 10% of them develop into chronic hepatitis,cirrhosis or HCC。The number of patients infected HCV is very large, the total number of which account for 40% of worldwide patient infected. Now the people more and more deeply understand the hepatitis virus, especially the development of hepatitis virus genomes. On the one hand ,the heterogeneity of the virus gene sequence knowed was studied,for example different genotype or subtype of HBV and HCV were sorted and identified,many drug-resistent variants or mutation loci were discovered and the conception of quasispecies was bringed forward. On the other hand,new hepatitis viruses were discovered, HGV,TTV and SENV were discovered in recent decade. With new research achievements coming forth,the disadvantages of clinically existing serum immunologic method and examination mostly using PCR technique are more and more obvious.Because they can not truly,roundly reflect all variances and dynamic changes of virus genome in vivo,the individual diagnosis and treatment was quite limited.In order to apply the gene research achievements of hepatitis virus during clincal practice,we need make use of new molecular biomedicine techniques to study and exploit new diagnostic methods and diagnostic productions of hepatitis virus. Recently,the biochip technique was widely used in a lot of fields.Because of its technologic advantages, including hepatitis virus,more than ten biochips of the pathogen or disease were reported worldwide.Hereby, we mainly study the preparation and application of HBV and HCV gene diagnostic microarray , as well as the experimental study of SENV, a new hepatitis virus.Experiment methods involved in this article include bioinformation analysis(namely the design of probe on chip and PCR primer,the search on web and comparation nucleotide sequence ),the synthezation of probes and primers,directly sequencing of PCR products,preparation method of probe on the microarray,the extraction of virus in serum,PCR amplification and digoxin label,hybridization and color development,data analysis of chip hybridization results and so on. the optimization program during chip preparation include: The selection of nylon membrane types and irradiation dosage when probes were fixed,the confirmation of the added-tail base numbers in probe′s 3′-terminal,the validation of chip hybridization using...