深圳地区HBV基因分型及耐药基因检测分析

Genotyping of HBV and detection of drug resistance gene in Shenzhen area

目的回顾分析深圳地区慢性乙型肝炎患者乙型肝炎病毒基因型别及耐药基因突变情况。方法采用实时荧光定量PCR法检测HBV DNA含量,免疫化学发光法检测HBe Ag含量,DNA反相斑点杂交芯片技术检测HBV DNA基因型与LMV和ADV常见耐药位点。结果 604例慢性乙型肝炎患者的病毒基因分型以B型为主,占62.91%;其次为C型,占31.13%。与B型相比,C型和B+C混合型的HBV DNA、HBe Ag及ALT水平均高于B型组。90例慢性乙型肝炎患者乙肝病毒基因出现不同位点耐药(90/604),耐药率为14.90%,其中阿德福韦耐药占所有耐药突变的62.22%(56/90),且以B型逆转录酶rt236T为主;拉米夫定耐药占所有耐药突变的37.78%(34/90),以B型逆转录酶rt108M为主,B+C混合型与其他型未见耐药基因突变型。结论深圳地区HBV基因型主要为B型,其次为C型;B、C型均可见对LMV和ADV耐药位点;基因芯片监测慢性乙型肝炎患者HBV基因型及耐药基因突变情况,有助于早期发现临床乙型肝炎耐药患者,指导临床合理用药。

Objective To investigate the genotype of HBV and drug resistance gene mutation in chronic hepatitis B patients in Shenzhen area. Methods The content of HBV DNA was measured by real-time fluorescence quantitative PCR, while the content of HBeAg was measured by chemiluminescence immunoassay, and DNA reverse Dot Blot hybridization chip technology was used to detected HBV genotype and drug-resistant mutant gene sites of lamivudine and adefovir. Results Type B hepatitis B virus accounted 62.91% of the 604 chronic hepatitis B patients followed by type C accounted for 31.13%. Compared with the type B, HBV DNA, HBeAg and ALT levels were much higher in type C and type B＋C. Drug resistance gene mutation sites were detected in 90 cases of HBV with the resistant rate of 14.90%. The resistance to adefovir accounted for 62.22% （56/90） of all drug resistance mutations and with type B reverse transcriptase rt236T predominated; lamivudine resistance was observed in type B reverse transcriptase rtl08M, accounted for 37.78% （34/90）; drug-resistant mutant gene sites were not noticed in type B ＋ C or other types. Conclusion Type B is the main genotype of HBV in Shenzhen, followed by type C ; type B and type C mutant genes are resistant to lamivudine and adefovir. Monitoring of HBVgenotype and gene mutation in chronic hepatitis B patients by using gene chip is helpful for early detection of drug resistance ,guiding clinical rational use of antiviral drugs.