抗凝状态不同的血液标本对实时荧光定量PCR法测定HBV-DNA结果的差异性研究

Research on the Differences between Blood Samples Anticoagulated Different of Real-time Fluorescent Quantitative PCR method for the Determination of HBV-DNA Results

目的:探讨用不同抗凝剂及不含抗凝剂的真空采血管采集的标本,对实时荧光定量(PCR)法检测乙型肝炎病毒HBV-DNA结果的差异,为临床选择HBV-DNA标本采集容器提供依据,也为临床医生分析不同标本类型的HBV-DNA结果提供参考。方法:用不同抗凝剂及不含抗凝剂的真空采血管采集标本,全部利用沉淀煮沸裂解法提取HBV核酸采用实时荧光定量PCR法检测HBV-DNA的含量。结果:用不同抗凝剂的真空采血管采集的血浆样本测定的HBV-DNA含量无显著性差异(P>0.05),但经EDTA-2K和肝素锂抗凝的血浆标本与不含抗凝剂的血清标本测定的HBV-DNA含量比较,结果显著降低(P<0.05)。共检测128份样本,其中105例三种真空采血管所采集标本的检测结果均有数值,此105例总相关系数(r)为0.916(P<0.01)。结论:不含抗凝剂的真空采血管采集的血清标本测定的HBV-DNA含量显著高于抗凝的血浆标本。

Objective: To investigate the use of anticoagulants and vacuum without anticoagulant blood samples were collected for real-time fluorescence quantitative( PCR) difference method for detection of hepatitis B virus HBV-DNA results,provide the basis for clinical selection of HBV-DNA specimen collection containers,different types of HBV-DNA specimens analysis results provide the reference for clinicians. Methods: With different anticoagulants and vacuum without anticoagulant blood specimen collection,Extraction of HBV nucleic acids by precipitation boiling cracking method. The content of HBV-DNA was detected by realtime fluorescent quantitative PCR. Results: There was no significant difference in HBV-DNA content between the plasma samples collected from the vacuum collecting vessels with different anticoagulants( P >0.05). However,the plasma levels of EDTA-2 K and heparin anticoagulant plasma samples were significantly lower compared with the serum levels of HBV-DNA without anticoagulant( P <0.05). A total of 128 samples were detected,of which 105 samples were collected by three kinds of vacuum blood vessels. The total correlation coefficient( R) of the 105 cases was 0.916( P <0.01). Conclusions: The content of HBV-DNA in serum samples without anticoagulant is significantly higher than that in anticoagulant plasma samples