Light Cycler实时监测PCR定量分析血清HBV DNA

Quantitation of Hepatitis B Virus DNA by Real-Time PCR Using LightCycler

目的 检验LightCycler实时监测PCR(real timedetectionPCR ,RTD PCR)对血清中HBVDNA定量检测的灵敏性和可重复性 ,探讨HBV血清标志物与HBVDNA定量的关系。方法 HBV定量按深圳匹基公司乙肝PCR荧光检测试剂盒使用说明 ,对乙肝标志物已明确的 773例血清中HBVDNA定量结果进行统计分析。结果 实时监测PCR对血清中HBVDNA定量检测的灵敏性高 ,可检测低至 1 0 0 0拷贝 /ml血清 ;可重复性好 ,批间误差 <2 0 % ;各种标志物类型的血清HBVDNA含量分布情况表明 ,HBV血清标志物中HBeAg与HBVDNA含量有明显的关系 ,一般HBeAg阳性血清HBVDNA含量较高 ,但也有相当一部分例外。结论 LightCycler实时监测PCR对血清中HBVDNA定量检测灵敏性高可重复性好 ;仅根据HBV血清标志物往往不能确定乙肝患者HBVDNA复制水平的高低 ,HBVDNA定量检测具有十分重要的临床意义。

Objective To test the sensitivity and repeatability of LightCycler real time PCR assay, and to explore the relationship between the nucleic acid content and serum markers of hepatitis B virus. Method LightCycler real time PCR assay was performed according to the operating instruction of PG Company HBV PCR detection kit. HBV DNA content and HBV serum markers of 773 patients were analyzed. Results The LightCycler real time PCR assay was sensitive and reproducible, which can detect 1000 copies of HBV DNA/ml sample with a deviation <20%. Serum HBV DNA content was related to the HBeAg serum markers. In general, HBeAg sero positive serum contained more HBV DNA, but with some exceptional cases. Conclusion LightCycler real time detection PCR assay is a sensitive and highly reproducible assay. Based only on the HBV serum markers, we could not estimate the DNA replication level. Therefore, quantitative measurement of the HBV DNA content by real time PCR is important in the clinical diagnosis.