摘要
目的:建立乙型肝炎病毒(hepatitis B virus,HBV)B基因型亚型的检测方法,并对其进行临床研究.方法:根据GenBank中20株Ba亚型,10株B j亚型以及25株C型全长基因组序列设计Ba、Bj亚型特异探针以及巢式PCR引物.采用反向杂交技术将B亚型特异探针固定在芯片上,通过与地高辛标记的扩增产物杂交检测B型HBV亚型.对镇江地区200例B基因型HBV血清样本进行亚型分析并对患者HBV DNA YMDD变异进行检测,通过对部分血清中的HBV DNA进行序列分析验证该方法的准确性.结果:200例B基因型HBV患者中,Ba亚型184例(92%),Bj亚型16例(8%).在持续使用拉米呋啶的80例患者中,Bj亚型6(0%)例,均无YMDD变异,Ba亚型74例,YMDD变异21(28.4%)例.结论:镇江地区B基因型HBV以Ba亚型为主,Ba、Bj亚型在服用拉米呋啶1年后发生YMDD变异的几率并无统计学上的显著差异.
AIM： To develop a new method of detecting subtypes of hepatitis B virus （HBV） genotype B and to investigate its clinical application and significance. METHODS： The probes specific to Ba and Bj subtypes of HBV were designed according to the phylogenetic analysis of 20 Ba, 10 Bj and 25 genotype C complete HBV genome sequences in GenBank. This method was based on the prin-ciple of reverse hybridization with Dig-labeled amplicons hybridizing to type-specific oligo-nucleotide probes that were immobilized on slides. Two hundred genotype B serum samples in Zhenjiang area were detected using the oli-gonucleotide chip, including 20 （ASC） and 180 （CH）. The results of randomly chosen sera were confirmed by direct sequencing.RESULTS： Of 200 HBV genotype B, subtype Ba and Bj accounted for 92% （n = 184） and 8% （n = 16）, respectively. Of 80 cases treated with Lami-vudine for 12 months, there were 6 Bj subtypes without YMDD mutations, and there were 74 cases of Ba subtypes with 21 YMDD mutations. CONCLUSION： Ba subtype prevails in Zhenji-ang area. The oligonucleotide chip is a reliable and convenient tool for the detection of HBV subtype. There is no significant difference of the response to the therapy between the two sub-types.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第27期3091-3094,共4页
World Chinese Journal of Digestology
基金
镇江市社会发展资助项目
No.SH2006046~~
