[1]苗丽,张秀平,王建昌,等.肉制品中鸡源性成分重组酶介导等温扩增检测方法的建立及应用[J].江苏农业学报,2019,(04):954-959.[doi:doi:10.3969/j.issn.1000-4440.2019.04.029]
　MIAO Li,ZHANG Xiu ping,WANG Jian chang,et al.Establishment and application of realtime recombinaseaided amplification assay to detect chickenderived ingredients in meat products[J].,2019,(04):954-959.[doi:doi:10.3969/j.issn.1000-4440.2019.04.029]
点击复制

肉制品中鸡源性成分重组酶介导等温扩增检测方法的建立及应用()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年04期
页码:
954-959
栏目:
加工贮藏·质量安全
出版日期:
2019-08-31

文章信息/Info

Title:
Establishment and application of realtime recombinaseaided amplification assay to detect chickenderived ingredients in meat products
作者:
苗丽1; 张秀平2; 王建昌3; 王永亮1; 程奇4; 徐超1; 张璐2
(1.郑州海关，河南郑州450003;2.河南检验检疫鉴定咨询中心，河南郑州450003;3.石家庄海关，河北石家庄050051;4.中国农业科学院生物技术研究所，北京100081)
Author(s):
MIAO Li1; ZHANG Xiuping2; WANG Jianchang3; WANG Yongliang1; CHENG Qi4; XU Chao1; ZHANG Lu2
(1.Zhengzhou Customs of the People′s Republic of China, Zhengzhou 450003, China;2.Identification Consulting Center of Henan Inspection and Quarantine, Zhengzhou 450003, China;3.Shijiazhuang Customs of the People′s Republic of China, Shijiazhuang 050051, China;4.Biotechnology Research Institute of the Chinese Academy of Agricultural Sciences, Beijing 100081, China)
关键词:
重组酶介导等温扩增; 细胞色素B基因; 鸡源性成分
Keywords:
recombinaseaided amplification; cytochrome B gene; chickenderived ingredients
分类号:
TS207
DOI:
doi:10.3969/j.issn.1000-4440.2019.04.029
文献标志码:
A
摘要:
为了快速准确检测出肉制品中的鸡源性成分，本研究基于鸡线粒体细胞色素B基因(CytB)，设计特异性引物和exo探针，建立了实时荧光重组酶介导等温扩增（RAA）方法，进行特异性、灵敏性和稳定性分析，并通过检测市售肉制品对所建立RAA方法的准确性和适用性进行分析。结果表明，该方法可以快速实现对高山鸡肉、乌鸡肉、芦花鸡肉、白羽鸡肉和野鸡肉基因组DNA的特异性扩增，而对猪肉、牛肉、羊肉等的DNA均没有扩增，可稳定检出的鸡源性成分最低质量分数为01%。在9份标识有鸡肉成分的市售样品中，7份检出鸡源性成分，有2份样品的鸡源性成分检测结果为阴性。本研究建立的实时荧光RAA方法操作简单，反应迅速，特异性强，灵敏度高，为肉制品中鸡源性成分的检测提供了技术保障。
Abstract:
In order to detect the Gallus ingredient in meat products quickly and accurately, specific primers and exo probes were designed based on chicken mitochondrial cytochrome B gene (CytB). A recombinaseaided amplification (RAA) realtime fluorescent method was established, and the specificity, sensitivity and stability were analyzed. Furthermore, the accuracy and applicability of RAA method were analyzed by testing meat products sold in the market. The results showed that this method could detect Gallus ingredient quickly. No crossreactions were found in specimens containing pork, beef and mutton. The detection limit was 01%. Detection results of commercially available samples showed that two samples were negative for chickenderived ingredients. The established method in this study has strong specificity and high sensitivity, providing a technical guarantee for the detection of chickenderived ingredients in meat products.

参考文献/References:

[1]ALI M E, HASHIM U, KASHIF M, et al. Development of swinespecific DNA markers for biosensorbased halal authentication[J]. Genetics and Molecular Research, 2012, 11(2): 1762-1772.
[2]KOPPEL R, DANIELS M, FELDERER N, et al. Multiplex realtime PCR for the detection and quantification of DNA from duck, goose, chicken, turkeyand pork[J]. European Food Research and Technology, 2013, 236（6）: 1093-1098.
[3]史艳宇,刘金华,吴月丹,等. 荧光定量PCR方法检测畜肉食品中鸭源性成分[J].食品安全质量检报, 2013,4(6):1859-1864.
[4]许如苏，周广彪，魏霜，等.锁核酸探针技术快速检测鸭源性成分的试验[J].中国兽医杂志，2015, 51（9）:91-93.
[5]FLOREN C, WIEDEMANN I, BRENIG B, et al. Species identification and quantification in meat and meat products using droplet digital PCR (ddPCR)[J]. Food Chemistry, 2015, 173: 1054-1058.
[6]刘岑杰，刘彦泓，杨滴，等. 肉制品中鸭源性成分的实时荧光PCR检测[J]. 肉类工业，2015（1）: 51-53.
[7]GILL P, GHAEMI A. Nucleic acid isothermal amplification technologies: a review[J]. Nucleosides, Nucleotides and Nucleic Acids, 2008, 27(3): 224-243.
[8]WALKER G T, LITTLE M C, NADEAU J G, et al. Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system[J]. Proceedings of the National Academy of Sciences, 1992, 89(1): 392-396.
[9]WALKER G T, FRAISER M S, SCHRAM J L, et al. Strand displacement amplification——an isothermal, in vitro DNA amplification technique[J]. Nucleic Acids Research, 1992, 20(7): 1691-1696.
[10]吕蓓,程海荣,严庆丰,等.体外核酸快速扩增技术的发展和不断创新[J]. 中国生物工程杂志, 2011, 31(3): 91-96.
[11]吕蓓,程海荣,严庆丰,等. 用重组酶介导扩增技术快速扩增核酸[J]. 中国科学, 2010, 40（10）:983-988.
[12]郭燕华,王德莲,王强,等. 重组酶介导等温扩增技术快速检测牛肉及牛肉制品中的牛源性成分[J]. 食品安全质量检测学报, 2017, 8(5): 1745-1749.
[13]KARABASANAVAR N S, SINGH S P, KUMAR D, et a1. Detection of pork adulteration by highlyspecific PCR assay of mitochondrial Dloop[J]. Food Chemistry, 2014, 145: 530-534.
[14]BALLIN N Z, VOGENSEN F K, KARLSSON A H. Species determinationCan we detect and quantify meat adulteration?[J]. Meat Science, 2009, 83(2): 165-174.
[15]张娟,宗卉,张利平. PCRmtDNA技术鉴别检测不同动物肌肉组织和饲料中鸭源性成分[J].生物工程学报, 2008, 24(10): 1832-1836.
[16]苗丽,张秀平,陈静,等.微滴数字PCR法定量检测肉制品中羊源性成分的研究[J].食品工业科技, 2016, 37(4):73-76.
[17]廖萍,刘茶珍,朱佩云,等. 重组酶介导扩增技术与传统聚合酶链反应技术在甲状腺癌DNA甲基化检测中的应用比较[J]. 中国医药, 2013, 8(6): 797-799.

相似文献/References:

[1]李大命,张彤晴,唐晟凯,等.太湖大银鱼(Protosalanx chinensis)细胞色素b基因序列多态性分析[J].江苏农业学报,2015,(04):840.[doi:10.3969/j.issn.1000-4440.2015.04.021]
　LI Da-ming,ZHANG Tong-qing,TANG Sheng-kai,et al.Genetic polymorphism of mitochondrial DNA cytochrome b (cytb) sequences of Protosalanx chinensis in Lake Taihu[J].,2015,(04):840.[doi:10.3969/j.issn.1000-4440.2015.04.021]

备注/Memo

备注/Memo:
收稿日期：2018-12-06 基金项目：河南省科技攻关项目（172102110202）；河北省科技计划项目（17275507D）；国家质检总局资助项目（2016IK114） 作者简介：苗丽（1971-），女，河南南阳人，硕士，高级兽医师，主要从事动物检疫及食品微生物学研究。（Tel）13523505628；(E-mail)ml5628@163.com
更新日期/Last Update: 2019-08-31