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当前位置: 首页 > 文章 > 草莓中甲型肝炎病毒检测 食品科学 2014,35 (24) 213-218
Position: Home > Articles > An Optimized Method for the Detection of Hapatitis A Virus(HAV) in Strawberry FOOD SCIENCE 2014,35 (24) 213-218

草莓中甲型肝炎病毒检测

作  者：
高世光;王海燕;郭慧;冯华炜;于洋;麻丽丹
单  位：
丹东出入境检验检疫局;大窑湾出入境检验检疫局
关键词：
草莓;甲型肝炎病毒;病毒富集;RNA提取
摘  要：
目的:建立草莓中甲型肝炎病毒(hapatitis A virus,HAV)的有效富集方法及病毒RNA提取方法,用于草莓中HAV检测。方法:利用甲肝减毒疫苗对已知阴性的草莓样品进行人工污染,通过有效富集条件的优化及RNA提取后,采用实时荧光逆转录-聚合酶链式反应进行检测。结果:病毒富集选择Tris-甘氨酸-1 g/100 m L牛肉浸提物缓冲液洗脱、果胶酶30 U、氯仿-正丁醇为抑制剂去除剂、聚乙二醇沉淀、5℃孵育1 h等优化条件,检测灵敏度较高;最优RNA提取方法为美国ABI公司生产的RNA提取试剂盒。采用优化后的方法对草莓样品中HAV病毒的检测显示,该病毒粒子的检测灵敏度可以达31.36 CCID50/20 g样品。同时对50份送检样品进行检测,结果均为阴性。结论:所建立的病毒富集方法和核酸提取方法更适合草莓样品中HAV的检测,灵敏度较高。
译  名：
An Optimized Method for the Detection of Hapatitis A Virus(HAV) in Strawberry
作  者：
GAO Shi-guang;WANG Hai-yan;GUO Hui;FENG Hua-wei;YU Yang;MA Li-dan;Dandong Entry-Exit Inspection and Quarantine Bureau;Dayaowan Entry-Exit Inspection and Quarantine Bureau;
关键词：
strawberry;;hepatitis A virus;;virus enrichment;;RNA extraction
摘  要：
Objective: To establish the optimal enrichment and RNA extraction procedures for detecting hepatitis A virus(HAV) in strawberries. Methods: After artificial contamination of known negative samples, HAV enrichment under optimized conditions and RNA extraction were performed for analysis by real-time fluorescence PCR. Results: The virus was enriched by elution with Tris glycine 1 g/100 m L beef extract buffer(TGBE), removal of the inhibitors using 30 U of pectinase and a mixture of chloroform with n-butanol, precipitation with PEG, and 1 h incubation at 5 ℃. The detection method was sensitive. The best RNA extraction results were achieved with a RNA extraction kit from ABI. The sensitivity of the optimized method for detecting HAV particles was 31.36 CCID50/20 g of sample. Negative results were reported for all 50 samples tested. Conclusion: The established method is suitable for the detection of HAV in strawberry samples with high sensitivity.

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