对丙型肝炎病毒核酸稳定性的研究

Study on Stability of Hepatitis C Virus RNA

目的探讨常规采血过程、标本采集及保存方式对HCV RNA稳定性的影响。方法采集HCV RNA阳性献血者的血样,以不同的抗凝剂、经不同的温度和时间保存后,采用荧光定量PCR方法测定HCV RNA病毒载量,考察HCV RNA的稳定性。结果不同抗凝剂下抗凝全血于4℃保存48h,HCV病毒载量未见显著降低(P>0.05);不同温度保存全血中,37℃组保存48h病毒载量显著降低(下降0.56Log);不同温度保存的血浆样品中,25℃保存7d,病毒载量出现显著降低(下降0.60Log);经反复冻融4次,血浆中病毒载量未见显著降低(P>0.05)。结论HCV较为稳定,常规血液采集、运输、实验室处理和保存等方式对其稳定性影响不大。

Objective To investigate whether processing and storage conditions could influence HCV RNA stability and therefore altered the detection Sensitivity for HCV NAT in whole blood or in plasma. Methods Samples from seven HCV RNA positive previous blood donors were kept in different storage conditions with different anticoagulants. HCV viral load was detected by fluorescent quantitative PCR method. Results There was no significant loss of HCV viral load in whole blood samples anticoagulated with CPDA,ACD,EDTA or none （P〉0.05）, while significant difference between the EDTA-anticoagulant storage condition with three other anticoagulants storage Conditions at 4℃ after 48 hours （P〈0.05）.The HCV viral load decreased by 0.27 Log, 0.14 Log and 0.56 Log after 48 hours of storage of whole blood anticoagulated with ACD at 4℃, 25℃ and 37℃ respectively. The HCV RNA level of plasma samples at 4℃ and at 25℃ （room temperature） after 7 days of storage decreased by 0. 15 Log and 0.60 Log respectively. After four freeze-thaw cycles the loss of HCV RNA was 0.41 Log in plasma samples. Conclusions HCV RNA is relatively resistant to degradation under routine blood collection, transport, laboratory handling and storage conditions.