标本处理方法对荧光定量PCR检测HBV DNA的影响

目的为提高标本检出率，比较煮沸法、Chelex 100树脂法和核酸纯化法等3种不同标本处理方法对荧光定量聚合酶链反应（FQPcR）技术检测乙型肝炎（简称乙肝）病毒（HBV）DNA的影响。方法收集36例乙肝确诊患者和14例非乙肝患者的血标本，分别经煮沸法、Chelex 100树脂法和核酸纯化法平行处理后，采用FQ-PCR进行检测。结果经3种方法处理的标本HBVDNA的检出率分别为52．78％、55．56％和100．00％。核酸纯化法的检出率明显高于其他2种方法（P〈0．01）。结论标本处理质量对进行HBVDNAFQ-PCR检测十分重要，煮沸法和Chelex 100树脂法不适用于临床标本检测，用核酸纯化法处理标本可保证FQ-PCR检测结果的准确性。

Effect of sample processing methods on detecting hepatitis B virus DNA by FQ-PCR

Objective The aim of the study was to compare the effect of three processing methods （boiling method,Chelex 100 resin method and nucleic acid purification method） on detecting HBV-DNA by FQ-PCR,and so as to improve the detection rate. Methods 36 patients with hepatitis B and 14 patients without hepatitis B were enrolled in the study,and blood samples were collected. Three different methods including boiling method, Chelex 100 resin method and nucleic acid purification method were applied to sample processing. FQ-PCR was employed to examine serum HBV-DNA. Results The detection rate of serum HBV-DNA was 52.78%, 55. 56% and 100% processed by boiling method,Chelex 100 resin method and nucleic acid purification method respectively. The nucleic acid purification method improved the detection rate significantly,prior to boiling method and Chelex 100 resin method（P 〈0.01）. Conclusion The quality of clinical sample preparation is vital to detecting hepatitis B virus DNA by FQ-PCR. The results suggested that the DNA templates prepared by boiling method and Chelex 100 resin method were not suitable for hepatitis DNA quantitative PCR detection,and nucleic acid purification method can ensure the aecura cy for FQ-PCR assay.