冻鸡肉中金黄色葡萄球菌LAMP快速检测方法的建立
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摘要
[目的]建立冻鸡肉中金黄色葡萄球菌的LAMP检测方法。[方法]根据金黄色葡萄球菌特异性耐热核酸酶基因序列设计2对引物,建立LAMP检测方法,并用于检测采购自贵阳市各大超市的冻鸡肉样品。[结果]设计合成的2对引物和建立的LAMP方法检测金黄色葡萄球菌具有良好的特异性,反应体系的灵敏度为65 CFU/m L。检测采购自贵阳市各大超市和经人为污染的264份冻鸡肉样品分别用LAMP方法检测,并用国标方法检测验证,其特异性为95.2%,准确度为97.3%。[结论]该研究建立的方法为实现在冻鸡肉中金黄色葡萄球菌的快速检测奠定了良好的基础。
[Objective] The research aimed to establish a LAMP method for detection of S. aureus in frozen chicken. [Method] Two pairs of primers were designed based on the nuc gene sequence of S. aureus. A LAMP detection method was established and used to detect frozen chicken samples from major supermarkets in Guiyang. [Result]Two pairs of primers and the established LAMP method had good specificity for the detection of S. aureus. The sensitivity was 65 CFU/m L. 264 frozen chicken samples purchased from major supermarkets in Guiyang and polluted by humans were detected by LAMP method and verified by national standard method. The specificity was 95. 2% and the accuracy was 97. 3%. [Conclusion]The method established in this study laid a good foundation for rapid detection of S. aureus in frozen chicken.
[Objective] The research aimed to establish a LAMP method for detection of S. aureus in frozen chicken. [Method] Two pairs of primers were designed based on the nuc gene sequence of S. aureus. A LAMP detection method was established and used to detect frozen chicken samples from major supermarkets in Guiyang. [Result]Two pairs of primers and the established LAMP method had good specificity for the detection of S. aureus. The sensitivity was 65 CFU/m L. 264 frozen chicken samples purchased from major supermarkets in Guiyang and polluted by humans were detected by LAMP method and verified by national standard method. The specificity was 95. 2% and the accuracy was 97. 3%. [Conclusion]The method established in this study laid a good foundation for rapid detection of S. aureus in frozen chicken.
引文
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[11]张嘉荟,方志宇,乔文姝.实时荧光定量PCR技术对金黄色葡萄球菌的检测概况[J].食品安全导刊,2017(27):111.
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[13]黄潇航,陈淑萍,李涵,等.基于LAMP技术的微生物病原诊断研究进展[J].产业与科技论坛,2018,17(1):52-53.
[14]ZHAO X,LI Y,PARK M,et al.Loop-mediated isothermal amplification assay targeting the fem A gene for rapid detection of Staphylococcus aureus from clinical and food samples[J].J Microbiol Biotechnol,2013,23(2):246-250.
[15]张艳艳,叶倩,王正荣,等.基于cox2基因的细粒棘球绦虫环介导等温扩增检测方法的初步建立[J].中国寄生虫学与寄生虫病杂志,2017(2):169-172.
[16]王利刚,张磊,张婧,等.国标培养法与2种金黄色葡萄球菌快速筛检方法的比较[J].食品安全质量检测学报,2017(1):238-242.
[17]WEI C J,ZHONG J L,HU T,et al.Simultaneous detection of Escherichia coli O157:H7,Staphylococcus aureus and Salmonella by multiplex PCR in milk[J].3 Biotech,2018,8(1):76.
[18]李琳,黄金海,赵耘,等.葡萄球菌肠毒素基因分型PCR检测技术的研究[J].食品科学,2008,29(7):340-344.
[2]GITAU G K,BUNDI R M,VANLEEUWEN J,et al.Evaluation of PetrifilmsTMas a diagnostic test to detect bovine mastitis organisms in Kenya[J].Trop Anim Health Prod,2013,45(3):883-886.
[3]顾其芳,张红芝,朱颖莹,等.三种金黄色葡萄球菌选择性分离培养基的检测效果比较[J].上海预防医学,2018(1):69-73.
[4]韩善桥,赵晓航,石峰,等.测试片法在金黄色葡萄球菌感染快速检测中的应用[J].国际检验医学杂志,2015,36(17):2459-2460,2463.
[5]孔祥瑞,王洪柱.金黄色葡萄球菌检测方法的研究进展[J].中国乳业,2017(10):72-74.
[6]杜志荣,高海宁,罗静.一起金黄色葡萄球菌引起的学校食物中毒事件调查分析[J].中国卫生产业,2015(12):20-22.
[7]张峰,顾敏霞,戴正,等.一起金黄色葡萄球菌肠毒素食物中毒事件调查[J].中国农村卫生事业管理,2017,37(7):803-805.
[8]王锦彤,田建伟,熊定凯.供列车冻鸡肉中同时检出肠炎沙门菌和金黄色葡萄球菌[J].中国国境卫生检疫杂志,2009(1):43-44.
[9]YAZDANKHAH S P,SLVERD L,SIMONSEN S,et al.Development and evaluation of an immunomagnetic separation-ELISA for the detection of Staphylococcus aureus thermostable nuclease in composite milk[J].Vet Microbiol,1999,67(2):113-125.
[10]董蕾,刘慧敏,孟璐,等.快速检测金黄色葡萄球菌活菌的研究进展[J].食品工业,2017(10):253-259.
[11]张嘉荟,方志宇,乔文姝.实时荧光定量PCR技术对金黄色葡萄球菌的检测概况[J].食品安全导刊,2017(27):111.
[12]NOTOMI T,OKAYAMA H,MASUBUCHI H,et al.Loop-mediated isothermal amplification of DNA[J].Nucleic Acids Res,2000,28(12):1-7.
[13]黄潇航,陈淑萍,李涵,等.基于LAMP技术的微生物病原诊断研究进展[J].产业与科技论坛,2018,17(1):52-53.
[14]ZHAO X,LI Y,PARK M,et al.Loop-mediated isothermal amplification assay targeting the fem A gene for rapid detection of Staphylococcus aureus from clinical and food samples[J].J Microbiol Biotechnol,2013,23(2):246-250.
[15]张艳艳,叶倩,王正荣,等.基于cox2基因的细粒棘球绦虫环介导等温扩增检测方法的初步建立[J].中国寄生虫学与寄生虫病杂志,2017(2):169-172.
[16]王利刚,张磊,张婧,等.国标培养法与2种金黄色葡萄球菌快速筛检方法的比较[J].食品安全质量检测学报,2017(1):238-242.
[17]WEI C J,ZHONG J L,HU T,et al.Simultaneous detection of Escherichia coli O157:H7,Staphylococcus aureus and Salmonella by multiplex PCR in milk[J].3 Biotech,2018,8(1):76.
[18]李琳,黄金海,赵耘,等.葡萄球菌肠毒素基因分型PCR检测技术的研究[J].食品科学,2008,29(7):340-344.