Study On The Preparation Of Gene Chip For HBV Genotype Analysis

Objective: To set up a method based on gene chip for determine HBV genotype, and putted it into practical use to investigate its value of diagnosis. Methods: the experimental condition for preparing the gene chips was studied and 7 fragments were screened from GenBank as probes dotted on chips to make HBV genotyping gene chips, many examinatorial experiments were done to evaluate the quality of this gene chip, and to analysis its detection characteristics. Results: hybridization temperature was defined at 39℃ based on lots of experiments, distinguished the test result by statistic software: fluorescence signal value above 30 was positive, or was negative. HBV-DNA negative serum samples and containing ordinary virus except HBV serum samples were tested by this chip, all of them was negative, 10 cases of clinical HBV-DNA positive serum samples test had the conclusion: 6 cases C type, 4 cases B type, and sequence analysis of 2 cases among them verified that gene chips were available. Standard serum of B type, C type were used to examine the gene chip: the result of standard serum test showed a coincidence rate of 100%, and the gene chip could test the minimum virus quantity was 104 copy/ml, gene chip had the total CV 9% and work function character for all tests was stable. Conclusion: HBV genotyping gene chip was sensitive and specific for HBV genotype determination, it can be used widely as a highly efficient, accurate HBV genotype detecting method for clinic diagnosis .