定量PCR检测血清中乙型肝炎病毒DNA及其意义

Quantitative detection of hepatitis B virus DNA in sera and its significance

目的检测血清中乙型肝炎病毒DNA拷贝数,并了解HBV感染的不同血清学指标组合相应的HBV DNA含量分布,以指导临床。方法 采用定量PCR和定性PCR方法,检测216份不同临床类型血清标本的HBV DNA,再用ELISA方法测定HBV-M,统计不同免疫指标组合的HBV DNA平均含量。结果病毒量分为高、中、低三度,大于107拷贝mL-1为高滴度:107-105拷贝mL-1为中等滴度;105拷贝mL-1以下为低滴度。60例HBsAg(+)HBeAg(+)HBcAb(+)血清,HBV DNA全部阳性,平均含量为1.9×108拷贝mL-1。51例HBsAg(+)HBeAb(+)HBcAb(+)血清,HBV DNA平均含量为5.4×106拷贝mL-1;33例HBsAg(+)HBeAb(+)血清,HBV DNA平均含量为7.5×105拷贝mL-1;114例HBsAb(+)HBeAb(+)HBeAb(+)血清,HBV DNA平均含量为1.8×105拷贝mL-1。定性和定量PCR阳生率分别为59.3％和61.6％。两种方法相对符合率为94.5％。结论定量PCR可真实反应HBV感染、复制及病程变化,对乙型肝炎临床诊断及治疗均有较大的指导意义。

Objective To detect the copy number of HBV DNA in sera and to know the amount of HBV DNA in different sere-logical marker combinations. Methods HBV DNA was detected by quantitative PCR, and HBV-M was detected by ELBA. The mean amount of HBV DNA was counted in different serological marker combinations. Results The quantity of HBV DNA was divided into high,middle and low liter.More than 107 copies mL-1 were the high liter.The middle liter was between 107 copies mL-1 and 105 copies mL-1. Less than 105 copies mL-1 was the low liter. In 60 HBsAg( + )HBeAg( + )HBcAb( + )samples,HBV DNA was positive, with 1.9 × 108 copies mL-1of HBV on average.In 51 HBsAg( + )HBeAb( + )HBcAb( + )samples,the average was 5.4 × 106 copies mL-1.In 33 HBsAg( + )HBcAb( + )samples,the average was 7.5 × 105 copies mL-1.In 14 HBsAb( + )HBeAb( + )HBcAb( + )samples,the amount was 1.8 × 105 copies mL-1. By quantitative and qualitative PCR, the positive rates were 61.6% and 59.3% , respectively.The results of both methods were very accordant. Conclusion Quantitative PCR can be used to monitor the true state of HBV replication and the course of disease.