贝类中甲肝病毒提取方法的比较

Comparison of extraction methods of hepatitis A virus in shellfish

中文关键词:  贝类  甲肝病毒  试剂盒  提取  比较

英文关键词:shellfish  hepatitis A virus  kits  extract  compare

中文摘要:

      目的 对贝类甲肝病毒的3种试剂盒提取方法进行比较。方法 在贝类样品中添加不同滴度的甲肝减毒疫苗后, 采用3种核酸提取法: Roche法、AM1836 法、Qiagen74104法。按照其说明书提取模拟样品病毒核酸模板, 从抽提效率、抽提核酸的稳定性、抑制剂去除效率3个方面对提取效果进行比较。结果 在抽提核酸稳定性、抑制剂去除效率方面, 3种方法结果相同, 稳定性的Ct值为24.795~28.651, 抑制剂去除效率方面提取的核酸10倍稀释核酸Ct值比未稀释核酸Ct值差值均为2.9～3.2左右(F=26.802, P<0.05); 在提取效率方面, Roche法高于AM1836法和Qiagen74104法, Ct值分别为28.5, 28.7和29.0。结论 3种方法中, Roche法在贝类中甲肝病毒提取方面更有优势, 可以提高贝类中甲肝病毒检测的灵敏度。

英文摘要:

      Objective To compare 3 extraction kits methods for hepatitis A virus in shellfish. Methods After adding different titers of attenuated hepatitis A vaccine to shellfish samples, 3 methods of nucleic acid extraction were used, including Roche method, AM1836 method and Qiagen74104 method. According to the instructions, the viral nucleic acid template of the simulated sample was extracted, and the extraction efficiency, the stability of the extracted nucleic acid and the removal efficiency of the inhibitor were compared. Results The stability of nucleic acid extraction and the removal efficiency of inhibitors of 3 methods were the same. The Ct value of the stability was 24.795-28.651. The difference of Ct values of 10-fold diluted nucleic acid and undiluted nucleic acid was about 2.9-3.2 (F=26.802, P<0.05), and the Roche method was superior to AM1836 and Qiagen74104 in extraction efficiency, and the Ct values were 28.5, 28.7 and 29.0 respectively. Conclusion Roche method has more advantages among the 3 methods in the extraction of HAV in shellfish, which can improve the detection sensitivity of HAV in shellfish.