终点法荧光定量PCR检测HBV-DNA的临床应用

目的运用终点法荧光定量PCR检测HBV-DNA,探讨其临床应用价值。方法运用本研究组制备的引物和荧光探针分别将HBV-DNA质粒及临床标本进行普通PCR和实时荧光定量PCR反应,再将普通PCR产物在荧光测定仪上测定荧光强度,与实时荧光定量PCR结果相比较,观察其符合程度。另外,运用相同方法比对本研究制备的PCR体系与市售荧光定量PCR试剂检测HBV-DNA的符合程度。结果终点法荧光定量PCR与实时荧光定量PCR检测HBV-DNA质粒及临床标本结果一致(P>0.05),与市售荧光定量PCR试剂比较,其符合程度较高(P>0.05)。结论终点法荧光定量PCR检测临床HBV-DNA标本稳定可靠,可作为荧光定量PCR的应用方法之一在基层医院推广应用。

Clinical application of endpoint detection of HBV-DNA by fluorescent quantitive PCR

Objective To investigate the clinical value of endpoint detection of HBV-DNA by means of fluorescent quantitive PCR.Methods The HBV-DNA plasmid and clinical specimens were respectively detected by ordinary PCR and fluorescent quantitive PCR through special primers and fluorescent probe which synthesised by our study. The fluorescent intensity of ordinary PCR products was detected by fluorescent analyzer, then compared with results of real time fluorescent quantitive PCR and estimated their coincidence degree subsequently. By the other hand, we matched our special FQ-PCR system against market sold FQ-PCR reagent kits through similar method and observed their coincidence degree.Results Results of HBV-DNA plasmid and clinical specimens which detected by endpoint FQ-PCR and real time FQ-PCR were all absolutely coincidence (P>0.05). There was no significant difference between our special FQ-PCR system and market sold FQ-PCR reagent kits on detecting HBV-DNA (P>0.05).Conclusion Endpoint fluorescent quantitive PCR is a stable and reliable method on clinical HBV-DNA detecting. It can be widespread application in clinical laboratories as a replacement method of real time FQ-PCR.