荧光定量分型PCR检测重庆地区乙型肝炎病毒感染者HBV基因型分布特征

Determination of distribution of hepatitis B virus genotype in infected population in Chongqing region by real-time fluorescent genotyping and quantitative PCR

目的了解重庆地区不同类型乙型肝炎病毒(HBV)感染者中HBV基因型的分布特点及其临床意义。方法应用荧光定量分型PCR(GQ-PCR)法检测207例HBV感染者的HBV基因型,并用直接测序法验证其基因分型结果的准确性。结果GQ-PCR成功分型204份临床样本,3例(1.45%)未能检出,其中,B型127例,占61.35%;C型29例,占14.01%;B/C混合型48例,占23.19%。C基因型的病毒载量明显高于B基因型及B/C混合基因型(P<0.05),B基因型与B/C混合基因型的病毒载量差异无统计学意义(P>0.05)。随机选取的90份血清样本经GQ-PCR和直接测序法检测其基因型,两种方法的检测结果一致性较好(Kappa=0.834,P<0.05)。结论重庆地区HBV感染者以B型为主要基因型,其次为B/C混合型,混合感染率高于以往研究结果。C基因型患者HBV-DNA水平高于B基因型及B/C混合基因型,可能与C基因型HBV感染导致肝脏损害较重有关。

Objective To investigate the distribution character of hepatitis B virus （HBV） genotype in infected population in Chongqing region by real-time fluorescent genotyping and quantitative PCR as well as its clinical significance. Methods The HBV genotypes of 207 infected subjects were determined by real-time genotyping and quantitative PCR （GQ-PCR）, and the results were verified by direct sequencing. Results Of the 207 samples, 204 were successfully genotyped, while 3 were not. Of the 204 samples, 127 （61. 35%） were judged as genotype B, 29（14. 01%） as genotype C, 48（23. 19%） as genotype B/C. The virus loads of samples of genotype C were significantly higher than those of genotype B and B/C （P 〈 0. 05 ）. However, no significant difference was observed between the virus loads of samples of genotypes B and B/C （P 〉 0. 05）. A total of 90 samples were randomly selected and genotyped by GQ-PCR and direct sequencing respectively, and the results were highly in agreement （Kappa = 0. 834,P 〈 0. 05 ）. Conclusion Genotype B is prominent in Chongqing region, secondary with genotype B/C. The mixed infection rate was higher than those reported previously. The HBV-DNA level of patients of infected with genotype C was higher than those with genotype B and B / C, which might be associated with more serious liver lesion caused by genotype C.