PCR荧光法对乙型肝炎病毒基因分型的检测及临床意义

目的 用荧光聚合酶链反应(PCR)技术,对乙型肝炎(HBV)患者基因型进行检测,了解该地区HBV基因型分布及其与肝功能损害、病毒复制水平的关系.方法 采用荧光聚合酶链反应(PCR),检测乙型肝炎病毒基因型,通过荧光探针识别基因型特异性序列,采集分析荧光信号确定病毒基因型,并分别检测谷丙转氨酶(ALT)水平、乙型肝炎病毒e抗原(HBeAg)和HBV-DNA含量.结果 532例慢性HBV感染者中,HBV基因型以C型为.主占60.2％,其次为B型占31.6％,B、C混合型占6.0％,未分型占2.3％例;C基因型HBV-DNA水平和HBeAg阳性率分别为(6.41±1.15)lg拷贝/ml和91.3％,明显高于B型(5.88±1.30)lg拷贝/ml和83.3％,差异有统计学意义(P＜0.01)；C基因型患者的ALT(130.16±197.19)U/L高于B基因型(125.6±145.02)U/L,但差异无统计学意义.结论 甘肃地区HBV基因型以C型为主,B型次之,C基因型HBV-DNA水平显著高于B基因型,C基因型HBeAg阳性率较B基因型高,C基因型对肝脏损害较B基因型重,并与HBV载量及HBeAg系统具有相关性.

Detection and genotyping of hepatitis B virus with RT-PCR and its clinical significance

OBJECTIVE To study the relationship between HBV genotypes and impairment of liver function as well as the level of hepatitis B virus replication by PCR.METHODS RT-PCR was adopted to detect the HBV genotypes;fluorescence probe was used to identify sequences of genotypes,and fluorescence signal was sampled and analyzed to confirm the viral genotypes.Levels of ALT,HBeAg and HBV-DNA were determined.RESULTS Among the 532 patients with chronic HBV infection,60.2% were found of genotype C,31.6% were of genotype B,6.0% were of genotype B mixed with genotype C,and 2.3% were of no genotype.The serum level of HBV-DNA was(6.41±1.15)lg copies/mL and the positive rate of HBeAg of genotype C was 91.3%,which were significantly higher than those of genotype B,(5.88±1.30)lg copies/mL and 83.3%,respectively(P<0.01).The level of ALT was higher in genotype C patients(130.16±197.19)U/L than in genotype B patients(125.6±145.02)U/L,but the difference was not statistically significant.CONCLUSION Genotype C is the predominant genotype and genotype B is the second common in this area.The serum level of HBV DNA of genotype C is significantly higher than that of genotype B;the positive rate of HBeAg of genotype C is higher than that of genotype B.Genotype B causes more serious damages to liver than genotype B does,which is closely related to the load of HBV and the expression of HBeAg.