微滴式數(shù)字聚合酶鏈?zhǔn)椒磻?yīng)對香腸制品中雞、豬、牛源性成分的定量分析

劉立兵 陳敏娜 孫曉霞 張亦琴 付琦 錢云開 周巍 郭春海 王建昌

摘 要：為實(shí)現(xiàn)肉制品中動物源性成分的定量檢測，基于微滴式數(shù)字聚合酶鏈?zhǔn)椒磻?yīng)（droplet digital polymerase chain reaction，ddPCR），以雞的轉(zhuǎn)化生長因子β-3基因、豬的朊蛋白基因和牛的生長激素基因?yàn)榘谢?，建立香腸制品中雞、豬、牛源性成分的定量檢測方法。結(jié)果表明：所建立ddPCR方法特異性強(qiáng)，能特異性檢測相應(yīng)的雞、豬、牛源性成分;根據(jù)肉粉質(zhì)量（mg）與DNA質(zhì)量濃度（ng/μL）、DNA質(zhì)量濃度（ng/μL）與基因拷貝數(shù)濃度（copies/μL）之間的線性關(guān)系，得到雞肉粉、豬肉粉和牛肉粉質(zhì)量（x1）與基因拷貝數(shù)濃度（y2）之間的關(guān)系式為：雞：;豬：;牛：（n為稀釋倍數(shù)）;基于建立的ddPCR方法對64 份市售不同種類香腸樣品進(jìn)行檢測，在1 份原料標(biāo)識僅為“豬肉”的香腸中檢出雞源性成分含量為18.68%。本研究建立的ddPCR方法能夠?qū)崿F(xiàn)香腸中雞、豬、牛源性成分的準(zhǔn)確定量檢測，并可以此判斷“蓄意摻假”或“無意帶入”。

關(guān)鍵詞：微滴式數(shù)字聚合酶鏈?zhǔn)椒磻?yīng);雞、豬、牛源性成分;定量分析;肉類摻假

Abstract： A droplet digital polymerase chain reaction （ddPCR） assay based on the transforming growth factor beta 3 gene of chicken， the porcine prion protein gene and the bovine growth hormone gene as the target genes were developed for the quantitative detection of chicken-， porcine- and bovine-derived ingredients in sausage products. The results showed that the established assay was highly specific for the detection of chicken-， porcine- and bovine-derived ingredients， but not for other tested animal components. Based on the linear relationship of DNA concentration （ng/μL） with meat mass （mg） and the number of gene copies per unit volume （copies/μL）， the linear equations between meat mass （x1） and the number of gene copies （y2） were established as follows： ?for chicken; ?for pork; and ?for beef （where n is dilution factor）. The ddPCR assay was used for the quantitative detection of 64 samples of different kinds of comercially available sausages. It was found that one of these samples， labelled as containing only pork， contained 18.68% chicken-derived ingredients. The ddPCR assay could allow accurate and quantitative analysis of chicken-， porcine- and bovine-derived ingredients in sausage， and could be used to discriminate intentional and unintentional adulteration.

Keywords： droplet digital polymerase chain reaction; chicken-， porcine-， and bovine-derived ingredients; quantitative analysis; meat adulteration

DOI：10.7506/rlyj1001-8123-20200508-115

中圖分類號：TS207.3 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 文獻(xiàn)標(biāo)志碼：A 文章編號：1001-8123（2020）08-0051-06

引文格式：

劉立兵， 陳敏娜， 孫曉霞， 等. 微滴式數(shù)字聚合酶鏈?zhǔn)椒磻?yīng)對香腸制品中雞、豬、牛源性成分的定量分析[J]. 肉類研究， 2020， 34（8）： 51-56. DOI：10.7506/rlyj1001-8123-20200508-115. ? ?http：//www.rlyj.net.cn

LIU Libing， CHEN Minna， SUN Xiaoxia， et al. Quantitative analysis of chicken-， porcine- and bovine-derived ingredients in sausage products by droplet digital polymerase chain reaction[J]. Meat Research， 2020， 34（8）： 51-56. DOI：10.7506/rlyj1001-8123-20200508-115. ? ?http：//www.rlyj.net.cn