多重微滴式数字PCR定量检测市售核桃乳中核桃、大豆源性成分
发布时间：2018-08-09 13:13
【摘要】：目的:为市售核桃乳中核桃源及主要掺杂物种大豆两种源性成分建立准确、快速定量检测方法。方法:从植物组织(核桃、大豆)或是植物蛋白饮料(核桃乳饮料)提取核酸后,主要采用多重微滴式数字聚合酶链式反应(polymerase chain reaction,PCR)技术,测算单位质量核桃和大豆的目的基因拷贝数比值,建立基因拷贝数与质量之间的关系,进而利用该比例关系换算出植物蛋白饮料中核桃和大豆的投料量,以实现对植物蛋白饮料中植物源性成分的定量检测。结果:基于多重微滴式数字PCR定量检测市售产品中大豆和核桃源性组分的方法,引物探针特异性好,目标物种间无交叉反应;灵敏度高,核桃中掺杂大豆的质量检测限为0.5%,相对误差为5.6%。将单一源性5种不同质量下靶基因拷贝数之比与5种不同比例混合源性靶基因拷贝数之比通过重复3次检测,综合比较并拟合后得到单位质量下拷贝数(C_(大豆)/C_(核桃)=1.771 1)的换算比例值。在从商品超市中抽取的11份不同品牌的核桃乳中(仅含核桃一种植物源成分),多重微滴式数字PCR方法检测显示:11份样品均检出核桃源性成分,6份样品检出大豆源性成分,其中4份样品中大豆与核桃质量之比高于10%,判断存在掺杂使假;2份样品大豆与核桃质量之比低于0.2%,极低的检出量推断为工艺沾染。结论:采用多重微滴式数字PCR准确、快速的定量方法可作为鉴别核桃乳中掺杂使假问题的有效手段。
[Abstract]:Objective: to establish an accurate and rapid quantitative detection method for the two components of walnut and soybean in walnut milk. Methods: after extracting nucleic acid from plant tissue (walnut, soybean) or vegetable protein beverage (walnut milk drink), the technique of (polymerase chain reaction-PCR was mainly used. The ratio of target gene copy number per unit weight of walnut and soybean was calculated, and the relationship between gene copy number and quality was established, and then the feeding amount of walnut and soybean in vegetable protein beverage was converted by using this ratio. In order to realize the quantitative detection of phytogenic components in vegetable protein drinks. Results: based on the method of quantitative detection of soybean and walnut origin components in commercial products by multiplex micro-drop digital PCR, the primer probe was specific and there was no cross reaction among target species, and the sensitivity was high. The quality detection limit of soybean adulterated in walnut was 0.5 and the relative error was 5.6. The ratio of the copy number of target gene to the copy number of target gene of 5 different mass of single source and 5 different proportion of mixed target gene was detected by repeating 3 times. The conversion ratio of copy number per unit mass (C _ (soybean) / C _ (walnut) 1.7771 1) was obtained by comprehensive comparison and fitting. In 11 samples of walnut milk of different brands (containing only one plant ingredient of walnut) extracted from commodity supermarket, the multiplex micro-drop digital PCR method showed that 6 samples of walnut origin were detected in 6 samples from 11 samples. The ratio of soybean to walnut mass in 4 samples was higher than 10. The ratio of soybean to walnut mass in 2 samples was lower than 0.2, and the very low detection quantity was inferred as technological contamination. Conclusion: the multiple microdrop digital PCR is accurate and rapid quantitative method can be used to identify the adulteration problem in walnut milk.
【作者单位】： 湖北省食品质量安全监督检验研究院;
【基金】：湖北省食品质量安全监督检验研究院自主立项科研项目(ZZLX2016009)
【分类号】：O652.9;TS275.4
，
本文编号：2174181
[Abstract]:Objective: to establish an accurate and rapid quantitative detection method for the two components of walnut and soybean in walnut milk. Methods: after extracting nucleic acid from plant tissue (walnut, soybean) or vegetable protein beverage (walnut milk drink), the technique of (polymerase chain reaction-PCR was mainly used. The ratio of target gene copy number per unit weight of walnut and soybean was calculated, and the relationship between gene copy number and quality was established, and then the feeding amount of walnut and soybean in vegetable protein beverage was converted by using this ratio. In order to realize the quantitative detection of phytogenic components in vegetable protein drinks. Results: based on the method of quantitative detection of soybean and walnut origin components in commercial products by multiplex micro-drop digital PCR, the primer probe was specific and there was no cross reaction among target species, and the sensitivity was high. The quality detection limit of soybean adulterated in walnut was 0.5 and the relative error was 5.6. The ratio of the copy number of target gene to the copy number of target gene of 5 different mass of single source and 5 different proportion of mixed target gene was detected by repeating 3 times. The conversion ratio of copy number per unit mass (C _ (soybean) / C _ (walnut) 1.7771 1) was obtained by comprehensive comparison and fitting. In 11 samples of walnut milk of different brands (containing only one plant ingredient of walnut) extracted from commodity supermarket, the multiplex micro-drop digital PCR method showed that 6 samples of walnut origin were detected in 6 samples from 11 samples. The ratio of soybean to walnut mass in 4 samples was higher than 10. The ratio of soybean to walnut mass in 2 samples was lower than 0.2, and the very low detection quantity was inferred as technological contamination. Conclusion: the multiple microdrop digital PCR is accurate and rapid quantitative method can be used to identify the adulteration problem in walnut milk.
【作者单位】： 湖北省食品质量安全监督检验研究院;
【基金】：湖北省食品质量安全监督检验研究院自主立项科研项目(ZZLX2016009)
【分类号】：O652.9;TS275.4
，
本文编号：2174181
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