Competitive annealing mediated isothermal amplification (CAMP) for rapid and simple detection of Listeria monocytogenes in milk

doi:10.1016/j.foodcont.2020.107347

	Competitive annealing mediated isothermal amplification (CAMP) for rapid and simple detection of Listeria monocytogenes in milk
	Li, Wei 2; Mao, Rui 4; Yue, Xiqing 3; Wu, Junrui 3; Wu, Rina 3; Qiao, Yu 1; Peng, Qing 1; Shi, Bo 1; Luo, Yong 5; Chen, Xu 3
刊名	FOOD CONTROL
	2020-11-01
卷号	117 页码:7
关键词	Competitive annealing mediated isothermal amplification L.monocytogenes Milk
ISSN号	0956-7135
DOI	10.1016/j.foodcont.2020.107347
英文摘要	Listeria monocytogenes (L. monocytogenes ) is widespread in nature and considered an important foodborne pathogen. And it can cause listeriosis, which can lead to miscarriage, neonatal death, septicemia and meningitis. Therefore, the appropriate detection methods are needed to effectively control and prevent this pathogenic bacterium. Competitive annealing mediated isothermal amplification (CAMP) is a novel nucleic acid-based detection technology that amplifies DNA with high sensitivity and specificity under isothermal conditions. The aim of this study was to develop a CAMP assay for rapid and simple detection of L. monocytogenes in milk. In this system, a pair of inner primers was designed to specifically target hlyA gene of L. monocytogenes. For further optimization, loop primers and outer primers were designed to accelerate CAMP reaction. Compared with conventional PCR method, the CAMP assay has the same specificity and higher sensitivity. All 8 strains of L. monocytogenes yielded positive results using the CAMP assay and showed no cross reaction with 26 tested non L. monocytogenes strains. The detection limit of the CAMP assay was 1.0 x 10(2) CFU/mL for pure bacterial, which was 10-fold more sensitive than PCR. Additionally, the CAMP method with the enrichment step could reliably detect L. monocytogenes in artificially contaminated milk samples at concentration as low as 1 CFU/mL. These results indicate the developed CAMP is a potential useful method for rapid detection of L. monocytogenes in milk samples.
资助项目	National key RD project[2018YFD0501403] ; Youth Project of the Education Department of Liaoning Province, China[LSNQN201911]
WOS关键词	REAL-TIME PCR ; MULTIPLEX PCR ; FOOD ; SALMONELLA ; STORAGE
WOS研究方向	Food Science & Technology
语种	英语
出版者	ELSEVIER SCI LTD
WOS记录号	WOS:000541152600014
资助机构	National key RD project ; Youth Project of the Education Department of Liaoning Province, China
内容类型	期刊论文
源URL	[http://ir.ipe.ac.cn/handle/122111/41176]
专题	中国科学院过程工程研究所
通讯作者	Chen, Xu; Du, Yuguang
作者单位	1.Chinese Acad Agr Sci, Feed Res Inst, Beijing 100081, Peoples R China 2.Shenyang Agr Univ, Dept Publicat, Shenyang 110866, Peoples R China 3.Shenyang Agr Univ, Food Sci Coll, Shenyang 110866, Peoples R China 4.Chinese Acad Sci, Inst Proc Engn, Beijing 100190, Peoples R China 5.Dalian Univ Technol, Sch Pharmaceut Sci & Technol, Dalian 116024, Peoples R China
推荐引用方式 GB/T 7714	Li, Wei,Mao, Rui,Yue, Xiqing,et al. Competitive annealing mediated isothermal amplification (CAMP) for rapid and simple detection of Listeria monocytogenes in milk[J]. FOOD CONTROL,2020,117:7.
APA	Li, Wei.,Mao, Rui.,Yue, Xiqing.,Wu, Junrui.,Wu, Rina.,...&Du, Yuguang.(2020).Competitive annealing mediated isothermal amplification (CAMP) for rapid and simple detection of Listeria monocytogenes in milk.FOOD CONTROL,117,7.
MLA	Li, Wei,et al."Competitive annealing mediated isothermal amplification (CAMP) for rapid and simple detection of Listeria monocytogenes in milk".FOOD CONTROL 117(2020):7.