基因芯片技术检测西藏拉萨地区的乙型肝炎病毒基因型  

Detection of HBV genotypes in Lhasa by gene chip

作　　者：曹占良[1] 高英堂[2] 刘霜[3] 景丽[4] 吉宗 刘彤[4] 刘持佳[1] 杜智[4] 

机构地区：[1]武警医学院附属医院检验科,天津市300162 [2]天津市第三中心医院天津市人工细胞重点实验室,天津市300170 [3]天津市人民医院检验科,天津市300120 [4]天津市第三中心医院 [5]武警西藏总队医院检验科,西藏自治区拉萨市850000

出　　处：《世界华人消化杂志》2007年第33期3500-3506,共7页World Chinese Journal of Digestology

基　　金：天津市科委攻关项目;No.05YFSZSF02500;武警医学院院级科研基金项目;No.WY2004-15~~

摘　　要：目的:研究西藏拉萨地区乙型肝炎病毒基因型的分布与特点.方法:采集92份西藏拉萨地区乙型肝炎患者的血清,参照GenBank中HBV DNA序列设计寡核苷酸探针并制备HBV基因分型芯片,利用套式PCR扩增HBV S基因部分片段,结合基因芯片、DNA测序和BioEdit软件进行基因分型检测,并对其与乙肝标志物、DNA含量、性别和民族之间的关系进行分析.结果:在92例血清标本中,套式PCR检测73例HBV DNA阳性可进行基因分型检测.其中B型13例(17.8%),C型18例(24.7%),D型39例(53.4%)和B/D混合基因型3例(4.1%).统计学分析3种基因型分布在不同乙肝标志物阳性、不同DNA含量和不同性别之间无差异,但与民族存在统计学差异(x^2=7.179,P<0.05).B型以汉族为主(9/13),而C、D型以藏族为主(12/18、28/39).将基因芯片分型的B、C、D型和B/D混合型进行DNA序列分析,表明两种分型方法的结果完全一致.结论:PCR结合基因芯片技术可用于HBV基因分型.西藏拉萨地区HBV基因型包括B、C、D和B/D混合型,其中以D型为主.AIM： To investigate the distribution and characteristics of HBV genotypes in Lhasa. METHODS： A total of 92 serum samples were collected from chronic hepatitis patients in Lhasa infected with HBV. Oligonucleotide probes were designed according to the DNA sequences from GenBank and used to prepare the genotyping chip. The fragments of HBV S gene were amplified using nested PCR. The genotypes were identified by gene chip and then confirmed by sequencing. The phylogeny was analyzed with BioEdit software. The relationship with hepatitis markers, DNA load, gender and race was also analyzed. RESULTS： Among the 73 patients positive for HBV DNA, 13 （17.8%） were genotype B, 18 （24.7%） genotype C, 39 （53.4%） genotype D, and 3 （4.1%） mixed genotype B/D. The distribution of the genotypes was not associated with serum HBV markers, HBV DNA load or gender, but was significantly associated with nationality （X^2= 7.179, P 〈 0.05）. Genotype g （9/13） was mostly distributed in the Han region, and genotypes C （12/18） and D （28/39） in Tibet. In addition, we selected 7 cases of genotype B, 6 of genotype C, 16 of genotype D, and 3 of mixed genotype B/D, identified by HBV typing chip which were further confirmed by sequencing, and the results of these two methods were consistent. CONCLUSION： Gene chip technology can be used for HBV genotyping. HBV genotypes distributed in Lhasa include B, C, D and B/D, of which genotype D is the predominant strain.

关 键 词：乙型肝炎病毒 基因型 基因芯片 序列分析 

分 类 号：R450[医药卫生—治疗学]