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首页>《中国测试》期刊>本期导读>基于核酸检测的虾过敏源尺度物质研究

基于核酸检测的虾过敏源尺度物质研究

87    2019-09-29

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作者:李春1, 高运华1, 王治栋1, 黄文胜2

作者单位:1. 中国计量科学研究院, 北京 100029;
2. 中国检验检疫科学研究院, 北京 100176


关键词:核酸检测;虾;过敏源;尺度物质


摘要:

选择最常食用的海产对虾作为虾过敏源尺度物质的候选物,提取并纯化其基因组DNA,采取序列测定法对设定的目标基因成分进行定性鉴定。用数字PCR技术精确确定海虾16S基因、虾真核生物18S基因、胡萝卜真核生物18S基因的拷贝数,以胡萝卜基因组DNA为本底,重量法配置虾16S基因拷贝数/真核生物18S基因组拷贝数比值为1%。评定虾16S基因拷贝数/真核生物18S基因拷贝数比值的定值不确定度,扩展不确定度为0.09%。并利用数字PCR仪对配置的尺度物质进行特性量值验证,结果一致性良好。可为基于核酸检测的食物过敏源尺度物质的研制提供方法参考。


Study on reference materials of shrimp allergen based on nucleic acid detection
LI Chun1, GAO Yunhua1, WANG Zhidong1, HUANG Wensheng2
1. National Institute of Metrology, Beijing 100029, China;
2. Chinese Academy of Inspection and Quarantine, Beijing 100176, China
Abstract: The marine shrimp was selected as a candidate for shrimp allergen reference materials, and its genomic DNA was extracted and purified. The target gene components were identified qualitatively by sequencing. The copies of shrimp 16S gene, shrimp and carrot eukaryotic18S gene were accurately determined by digital PCR. Based on the genomic DNA of shrimp 16S gene, shrimp and carrot eukaryotic 18S gene, the ratio for copies of 16S gene/18S gene of eukaryotic certified by gravimetric method was 1%. The uncertainty of DNA reference materials for the ratio for copies of 16S gene/18S gene of eukaryotic was evaluated, and the expanded uncertainty was 0.09%. Digital PCR was used to validate the characteristic values of the standard substances, and the results were in good agreement, which provided a reference for the development of food allergens reference substances based on nucleic acid detection in China.
Keywords: nucleic acid detection;shrimp;allergen;reference materials
2019, 45(9):49-53  收稿日期: 2019-02-15;收到修改稿日期: 2019-03-31
基金项目: 国家质量研发重点专项项目(2017YFF0204604);国家质检公益行业科研专项(201510026);尺度物质平台项目(32-APT1802-12)
作者简介: 李春(1994-),男,安徽长丰县人,硕士研究生,专业方向为核酸定量分析
参考文献
[1] EHLERT A, HUPFER C, BUSCH U. Overview of detection methods of allergens in foods. Fleischwirtschaft, 2008, 88(1):89-91.
[2] 周煌, 杨安树, 佟平, 等. 食物过敏源尺度物质的研究进展[J]. 食品工业科技, 2012, 33(23):438-442
[3] 龚方, 房保海. 国内外食品过敏原标签管理现状与趋势[J]. 食品保险质量检测学报, 2012, 3(3):226-230
[4] OLIVER S, STEFAN V. Development of a real-time PCR and a sandwich ELISA for detection of potentially allergenic trace amounts of peanut (arachis hypogaea) in processed foods[J]. Journal of Agricultural and Food Chemistry, 2004, 52(12):3754-3760
[5] 王玮, 韩建勋, 袁飞, 等. 多重PCR同时检测常见8种食物过敏原[J]. 中国食品学报, 2011, 11(6):152-157
[6] SIEGEL M, MUTSCHLER A, BOERNSEN B, et al. Food matrix standards for the quantification of allergenic food ingredients using real-time PCR[J]. European food research and technology, 2013, 237(2):185-197
[7] 韩远龙, 吴志华, 闫飞, 等. 花生过敏原检测方法研究进展[J]. 食品科学, 2012, 33(13):305-308
[8] CHO C Y, NOWATZKE W, OLIVER K, et al. Multiplex detection of food allergens and gluten[J]. Analytical and bioanalytical chemistry, 2015, 407(14):4195-4206
[9] ELISA I, ANA J, NURIA P, et al. Real Time PCR to detect hazelnut allergen coding sequences in processed foods[J]. Food Chemistry, 2013, 138(2/3):1976-1981
[10] 梁君妮, 孙敏. 实时荧光PCR法检测食物中榛子过敏原成分[J]. 食品工业科技, 2011(1):293-295
[11] 陈家杰, 王海燕, 梁秋妮, 等. 两种PCR方法检食品中花生过敏原Arah1成分[J]. 食品研究与开发, 2011, 32(9):69-74
[12] FUTOSHI A, TAKANE O. PCR-based detection of allergenic mackerel ingredients in seafood[J]. Journal of genetics, 2004, 83(2):193-195
[13] 出口食品中过敏原成分检测方法 第10部分 实时荧光PCR方法检测虾蟹成分:SNT 1961.10-2013[S]. 北京:中国质检出版社, 2013.
[14] 出口食品中过敏原成分检测方法 第7部分 实时荧光PCR方法检测胡萝卜成分:SNT 1961.7-2013[S]. 北京:中国质检出版社, 2013.