新型HBV-DNA实时荧光定量PCR试剂盒的应用

目的:观察新型HBV-DNA荧光定量PCR试剂盒的临床应用价值。方法:采用荧光定量PCR检测法,用10份正常血清、10份HBV高滴度(106 IU/ml)血清和梯度稀释的HBV克隆质粒标准品,验证该试剂盒检测结果的重复性、特异性和灵敏度;并将该10份HBV高滴度(106 IU/ml)血清结果与同时用传统方法所测结果进行配对检验。结果:10份正常血清标本均无假阳性HBV-DNA检测结果,特异性极好;灵敏度可达到103 IU/ml,重复性和准确性两者之间差异无显著性(t=1.15,P>0.05)。结论:该方法特异性和灵敏度高,方便、快捷,可替代传统试剂盒用于HBV的临床检测和科学研究。

The Application of New Type Real-time PCR Fluore Scence Detection for Hepatitis B Virus DNA Kit

Objective To observe the value of the new type real-time PCR fluorescence detection for hepatitis B virus DNA （HBV-DNA）kit in clinic examination.Methods Ten normal sera samples and ten sera samples with high level （106 IU/ml）HBV-DNA,and HBV plasmid standards with 1：10 serial dilutions were used to investigate the specificity, repetition and sensitivity of the new type real-time PCR fluorescence detection kit,Then given a statistics analysis for the results of the ten sera samples with high level（106 IU/ml）HBV-DNA detected by the new type kid（A method）and the traditional type kit （B method）.Results No significant fluorescent signal was observed when 10 normal sera samples were amplified, and lowest limitation of this assay determined by 1：10 serial dilutions of HBV cloned plasmid was 1031U/mL ,There was no significant difference for HBV-DNA results in the two methods （t= 1.15,P〉0.05）.Conclusion The new type real-time PCR fluores- cence detection kit is highly specific and sensitive,which can be substituted the traditional type kit and applied in the scientific research of HBV as well as in clinic examination.