Multiplex LAMP method for the detection of pathogenic bacteria in eggs

ZHAO Jun; CHEN Ze-ping; LI Zhen; SONG Xiao-ning; LI Yu-feng

doi:10.13386/j.issn1002-0306.2016.04.012

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Article Navigation > Science and Technology of Food Industry > 2016 > (04): 107-110

ZHAO Jun, CHEN Ze-ping, LI Zhen, SONG Xiao-ning, LI Yu-feng. Multiplex LAMP method for the detection of pathogenic bacteria in eggs[J]. Science and Technology of Food Industry, 2016, (04): 107-110. doi: 10.13386/j.issn1002-0306.2016.04.012

Citation:

ZHAO Jun, CHEN Ze-ping, LI Zhen, SONG Xiao-ning, LI Yu-feng. Multiplex LAMP method for the detection of pathogenic bacteria in eggs[J]. Science and Technology of Food Industry, 2016, (04): 107-110. doi: 10.13386/j.issn1002-0306.2016.04.012

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Multiplex LAMP method for the detection of pathogenic bacteria in eggs

doi: 10.13386/j.issn1002-0306.2016.04.012

College of Food and Biological Engineering,Xihua University

Received Date: 2015-07-17

Abstract

Abstract

Two strains of bacteria named X_1,X_2 were found to detect the pathogenic bacteria on eggs. Combined with 16 S r RNA molecule methods,testing the cloned plasmid sequence,blasting the sequence with NCBI in Gen Bank. The results showed that X_1 was Bacillus cereus NC7401(AP007209) and X_1 was Staphylococcus aureus(AP003088). According to the highly conservative gene nhe and nuc,two sets of primers were designed.The optimized LAMP reaction system contained Mg SO41.5 μL,each primer 2.0 μL and reaction temperature of60 ℃. The specificity was verified by using 9 strains of non-targeting bacteria and targeting bacteria X_1,X_2.The LAMP assay showed a sensitivity of 10 fg / μL. The novel multiple LAMP method reported here was characteristics of rapid detection, high specificity and excellent sensitivity, which could be applied in the detection of pathogenic bacteria.
- egg,
- pathogenic bacteria,
- 16S r RNA,
- loop-mediated isothermal amplification,
- rapid detection

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References(23)

References

[1]	滕仁明.食源性疾病[J].中国社区医师,2007,14(2):39-45.
[2]	王振国,刘金华,肖成蕊,等.利用PCR技术检测致病性蜡样芽孢杆菌的研究[J].生物技术,2005,15(5):45-47.
[3]	秦成峰,秦鄂德.环介导的恒温扩增技术及其在病毒性疾病诊断中的应用[J].生物技术通讯,2007,18(5):87-90.
[4]	东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001(2):21-53.
[5]	陈建辉,杨劲松,陈爱萍,等.应用PCR技术快速检测沙门菌[J].海峡预防医学杂志,2009,15(3):55-57.
[6]	王韶,刘桂华,乔凤,等.食品中金黄色葡萄球菌的PCR检测[J].吉林大学学报:医学版,2006,32(5):933-936.
[7]	K IM H J,CHO J C.Rapid and sensitive detection of Listeria monocytogenes using a PCR-Enzymelinked immunosorbent assay[J].Journal of Microbiology and Biotechnology,2008,18(11):1858-1861.
[8]	闫冰,姜毓君,曲妍妍,等.实时RT-PCR检测存活于乳中的单核细胞增多性李斯特菌[J].食品科学,2008,29(2):292-296.
[9]	SN/T 1869—2007食品中多种致病菌快速检测方法PCR法[S].
[10]	Nicola Pusterla a,Barbara A.Use of quantitative real-time PCR for the detection of Salmonella spp.in fecal samples from horses at a veterinary teaching hospital[J].The Veterinary Journal,2010,186:252-255.
[11]	万红梅,李玉锋,李帅.环介导等温基因扩增技术快速检测不耐热型肠毒素大肠杆菌反应条件优化[J].现代预防医学,2011,38(2):136-139.
[12]	涂宏钢,李存瑞,巫庆华,等.利用16S r RNA全长序列鉴定植物乳杆菌[J].乳业科学与技术,2004,2(2):57-60.
[13]	付晓艳,霍贵成.利用16S r RNA部分序列聚类分析鉴定乳球菌[J].中国乳品工业,2005,33(8):7-9.
[14]	乌日娜,张和平,孟和毕力格.酸马奶中乳杆菌ZLl2-1的16Sr RNA基因序列及聚类分析[J].中国乳品工业,2005,33(6):4-9.
[15]	周帼萍,袁志明.蜡样芽孢杆菌污染及对食品安全的影响[J].食品科学,2007,28(3):357-361.
[16]	唐梦君,周生,张小燕,等.应用LAMP快速检测金黄色葡萄球菌的研究[J].现代食品科技,2011,27(6):719-722.
[17]	易海华,祝长青,宋阳威,等.食品中单增李斯特菌环介导等温扩增检测技术[J].食品科学,2011,32(4):203-208.
[18]	姜侃,吕沁风,汪新,等.三重LAMP法检测食品中沙门氏、单增李斯特菌和金黄色葡萄球菌[J].食品科学,2013,34(24):182-187.
[19]	马寅众,陈江源,房国梁,等.环介导等温扩增法快速检测阪崎肠杆菌[J].食品科学,2010,31(22):322-326.
[20]	鲁玉侠,郭祀远,石磊,等.EMA-LAMP方法快速检测死/活的食源性沙门氏菌[J].食品科学,2009,30(22):324-328.
[21]	邵碧英,陈文炳,曾莹,等.LAMP法检测转基因大豆A2704-12品系[J].食品科学,2013,34(24):202-207.
[22]	程晋霞,周熙诚,马丹,等.食品中芥末过敏原成分LAMP检测方法的建立[J].食品科学,2014,35(20):148-153.
[23]	刘昊,黄文胜,邓婷婷,等.LAMP法检测食品中开心果过敏原成分[J].食品科学,2013,34(22):128-132.