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AIM: To determine the genotype distribution of hepatitis B virus (HBV) with a newly oligonucleotide chip assay among the HBV carriers in East China.METHODS: An assay using oligonucleotide chip was developed for detection of HBV genotypes in serum samples from HBV DNA-positive patients in East China. This method is based on the principle of reverse hybridization with Cy5-labeled amplicons hybridizing to type-specific oligonucleotide probes that are immobilized on slides. The results of 80 randomly chosen sera were confirmed by direct sequencing.RESULTS: HBV genotype B, C and mixed genotype were detected in 400 serum samples, accounting for8.3% (n = 33), 83.2% (n = 333), and 8.5% (n = 34),respectively. The evaluation of the oligonucleotide assay showed 100% concordance with the amplicon phylogenetic analysis except 9 mixed genotype infections undetected by sequencing.CONCLUSION: The study indicates that HBV genotype C and B prevail in the East China. It is suggested that the oligonucleotide chip is a reliable and convenient tool for the detection of HBV genotyping.