title: Genotyping of hepatitis B virus (HBV) by oligonucleotides microarray.
abstract: Oligonuleotides (oligo) microarray is a promising method for virus genotyping. In this study, we developed an oligo microarray used for genotyping hepatitis B virus (HBV). It consists of 15 probes targeting HBV genotypes A-G and two genotypes of apes, and one conserved probe selected from the HBV pre-S region. To test a clinical sample, the gene targets of clinical samples were amplified and labelled with Cy5-dCTP in a PCR reaction using primers covering the flanking region of the HBV pre-S gene. Following purification, the labelled PCR products were hybridized to the microarray. In an analysis of 96 HBV patients' serum samples using our developed microarray, we identified 34 genotype B, 60 genotype C and 2 genotypes B/C coinfection samples. Sequencing results of 24 randomly selected samples agreed 100% with microarray data. Geographic distribution of genotypes C and B was also divergent, 95.7% (44/46) of genotype C and 64.0% (32/50) of genotype B samples were collected from Northern and Southern China, respectively. The accuracy of genotyping was confirmed by analyzing DNA sequences of 24 samples. It is demonstrated that low-density oligo microarray can serve as a reliable and time-saving method to HBV genotyping from patient's sera.
authors.affiliation: Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Fengtai District, Beijing, People's Republic of China.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Song
authors.forename: Erhei
authors.initials: E
authors.lastname: Dai
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.forename: Hengjun
authors.initials: H
authors.lastname: Liu
authors.forename: Junhui
authors.initials: J
authors.lastname: Zhai
authors.forename: Cuiying
authors.initials: C
authors.lastname: Chen
authors.forename: Zongmin
authors.initials: Z
authors.lastname: Du
authors.forename: Zhaobiao
authors.initials: Z
authors.lastname: Guo
authors.forename: Ruifu
authors.initials: R
authors.lastname: Yang
mesh_terms: D016679:Genome, Viral; D005838:Genotype; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis