北京市市售贝类、蔬菜、浆果、即食海产品中诺如病毒污染状况检测及检测方法探析
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(1.中国食品药品检定研究院,北京 100050;2.吉林省长春市食品药品检验中心,吉林 长春 130012;3.甘肃省食品检验研究院,甘肃 兰州 730030)

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骆海朋 男 副主任检验技师 研究方向为食品微生物学 E-mail:haipengluo123@sina.com通信作者:┣┣(中)通信作者┫┫崔生辉 男 研究员 研究方向为食品微生物学 E-mail:cuishenghui@aliyun.com

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Survey on the contamination of Norovirus in shellfish, vegetable, fruit and ready-to-eat seafood from markets in Beijing
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(1.National Institute of Food and Drug Control,Beijing 100050,China;2.Jilin Province Inspection Center for Food and Drug,Jilin Changchun 130012,China;3.Gansu Province Food Inspection Institute,Gansu Lanzhou 730030,China)

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    摘要:

    对北京市市售贝类(牡蛎、贻贝、扇贝、文蛤、毛蚶)、蔬菜(苗菜、生菜)、浆果(草莓、蓝莓)、即食海产品(三文鱼、即食虾)进行诺如病毒检测。方法 贝类、蔬菜、浆果类样品参照ISO/TS 15216-1方法进行前处理,虾类检测参照贝类、三文鱼参照蔬菜的方法并进行改良。病毒RNA提取参照罗氏High pure viral RNA Kit试剂盒方法对样品前处理后的提取液进行提取。实时荧光逆转录聚合酶链反应(实时荧光RT-PCR)的检测使用罗氏LightMix noroviurus GI-GII于Lightcycler 480 II进行检测,并进行GI-GII分型。诺如病毒阳性样品,参照SN/T 2626—2010《国境口岸诺如病毒检测方法》,使用JV12、JV13进行扩增,将获得的DNA扩增片段进行测序并分型。结果 经罗氏试剂盒检测贝类食品478份(40份阳性)、蔬菜62份(1份阳性)、浆果84份(0份阳性)和即食海产品51份(1份阳性)。有42份为GI-GII,其中37份为GII型,4份为GI型。在42份阳性样品中,有29份GII型阳性样品对RNA聚合酶区域扩增成功,通过序列分析,27份为GII.17,1份为GII.12,1份为Hawaii.calicivirus。结论 通过对北京市市售食品样品中诺如病毒检测发现,贝类食品呈现较明显的季节分布,在冬季检出率较高,贝类食品是诺如病毒污染率最高的食品。

    Abstract:

    To investigate the contamination of Norovirus in shellfish (oysters, mussels, scallops, clams, clam), vegetables (seedling vegetables, lettuce), berries (strawberries, blueberries), ready-to-eat seafood (salmon, shrimp ) from markets in Beijing.Methods Pretreatment methods for shellfish, vegetables and berries samples were referred to ISO/TS 15216-1; pretreatment method for shrimp and shellfish was referred to shellfish method; pretreatment method for salmon was referred to improved vegetables method. Viral RNA extraction was referred to Roche High pure viral RNA Kit K methods. Real-time reverse transcription-polymerase chain reaction testing and GI-GII typing were carried out by Roche LightMix noroviurus GI-GII in Lightcycler 480 II; Norovirus positive samples was amplified using the prime of JV12, JV13 and the amplified DNA fragments were sequenced for subtyping. Results 478 shellfish (40 positive), 62 vegetables (1 positive), 84 berries (0 positive) and 51 raw salmon (1 positive) were detected. 35 positive samples were successfully amplified by sequence analysis. 27 samples were GII.17,1 sample was GII.12, and 1 sample was Hawaii.calicivirus.Conclusion Through the norovirus monitoring in Beijing market, shellfish presented obvious seasonal distribution and higher detection rate which suggested that the shellfish was the main sources of Norovirus in food.

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骆海朋,高飞,于海瑶,仝伟健,任秀,余文,崔生辉.北京市市售贝类、蔬菜、浆果、即食海产品中诺如病毒污染状况检测及检测方法探析[J].中国食品卫生杂志,2017,29(2):218-222.

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  • 收稿日期:2017-02-15
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  • 在线发布日期: 2017-05-03
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