食品科学 ›› 2019, Vol. 40 ›› Issue (6): 325-330.doi: 10.7506/spkx1002-6630-20180111-131

• 安全检测 • 上一篇    下一篇

食品中甲型肝炎病毒RNA提取方法的比较及应用

冯华炜1,闫平平2,艾海新1,3,李思嘉4,麻丽丹5,刘宏生1,3,*   

  1. 1.辽宁大学生命科学院,辽宁 沈阳 110036;2.大连出入境检验检疫局,辽宁 大连 116400;3.辽宁省生物大分子模拟计算与信息处理工程研究中心,辽宁 沈阳 110036;4.东港市农业综合执法大队,辽宁 丹东 118000;5.丹东出入境检验检疫局,辽宁 丹东 118000
  • 出版日期:2019-03-25 发布日期:2019-04-02
  • 基金资助:
    国家质检总局科研项目(2015IK167);国家自然科学基金面上项目(31570160);沈阳市科技局项目(17-231-1-04)

Comparison and Application of RNA Extraction Methods for Detection of Hepatitis A Virus in Food Samples

FENG Huawei1, YAN Pingping2, AI Haixin1,3, LI Sijia4, MA Lidan5, LIU Hongsheng1,3,*   

  1. 1. School of Life Sciences, Liaoning University, Shenyang 110036, China; 2. Danlian Entry-Exit Inspection and Quarantine Bureau, Dalian 116400, China; 3. Research Center for Computer Simulating and Information Processing of Bio-macromolecules of Liaoning, Shenyang 110036, China; 4. Donggang Comprehensive Agricultural Law Enforcement Brigade, Dandong 118000, China; 5. Dandong Entry-Exit Inspection and Quarantine Bureau, Dandong 118000, China
  • Online:2019-03-25 Published:2019-04-02

摘要: 目的:对食品中甲型肝炎病毒的3 种RNA提取方法进行综合比较,以优选出最佳的核酸提取方法,为各级实验室开展食源性甲型肝炎病毒核酸检验提供技术参考。方法:分别采用ABI AM1836、QIAGEN 74104和 ROCHE11858882001三种商业化核酸提取试剂盒对人工污染甲型肝炎病毒的食品样品进行核酸提取,根据3 种核酸提取方法的提取效率、抑制剂去除效率、检测灵敏度、综合应用指标进行病毒RNA的优化,且采用优化后的提取方法进行实际样品的检测。结果:综合病毒各项评价指标显示,ROCHE 11858882001在食品中提取甲型肝炎病毒RNA效果最优,其次为QIAGEN 74104和ABI AM1836。101 份样品的检测结果显示,一份蓝靛果样品为甲型肝炎病毒核酸阳性,其余样品均为阴性。结论:ROCHE 11858882001是一种快捷有效的病毒RNA提取方法,适用于食品中甲型肝炎病毒的日常检测工作。

关键词: 甲型肝炎病毒, RNA提取方法, 实时荧光RT-PCR, 食品样品

Abstract: Objective: Three RNA extraction kits were compared to select the best one for RNA extraction from hepatitis A virus (HAV) in foods, aiming to provide a technical basis for foodborne HAV RNA detection in laboratories. Methods: After artificial contamination of known negative samples, three commercial nucleic acid extraction kits, ABI AM1836, QIAGEN 74104 and ROCHE 11858882001, were used to extract HAV RNA and their performance was evaluated in terms of extraction efficiency, inhibitor removal efficiency and sensitivity, and the optimized extraction kit was applied to real samples. Results: ROCHE 11858882001 was the best extraction kit, followed by QIAGEN 74104 and ABI AM1836. Of 101 samples, one sample of blue honeysuckle was positive while the rest were negative. Conclusion: ROCHE 11858882001 is a rapid and effective method for extracting HAV RNA, and thus it is suitable for the routine detection of foodborne HAV.

Key words: hepatitis A virus, RNA extraction method, real-time fluorescence RT-PCR, food samples

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