Development of a real-time loop-mediated isothermal amplification assay for the sensitive and rapid detection of Listeria monocytogenes

被引：12
作者：
Ye, L. [1 ]
Li, Y. [1 ]
Zhao, J. [2 ]
Zhang, Z. [3 ]
Meng, H. [1 ]
Yan, H. [1 ]
Miyoshi, S. -i. [4 ]
Shi, L. [1 ]
机构：
[1] S China Univ Technol, Coll Light Ind & Food Sci, Guangzhou 510640, Guangdong, Peoples R China
[2] Guangdong Univ Petrochem Technol, Coll Environm & Biol Engn, Maoming, Peoples R China
[3] Xiamen Yinxiang Grp Co Ltd, State Key Lab Food Safety Technol Meat Prod, Xiamen, Peoples R China
[4] Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Tsushima, Okayama, Japan
来源：
LETTERS IN APPLIED MICROBIOLOGY | 2015年 / 61卷 / 01期
关键词：
API Listeria; ESEQuant tube scanner; Listeria monocytogenes; RealAmp; real-time PCR; VIBRIO-PARAHAEMOLYTICUS; PCR; IDENTIFICATION; STRAINS; SYSTEM; FOOD;
D O I：
10.1111/lam.12429
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
A real-time loop-mediated isothermal amplification (RealAmp) assay for the detection of Listeria was developed. The RealAmp assay, using primers specific for the hemolysin-encoding hlyA gene, was verified using Listeria monocytogenes strains (n=58) from different regions of the world. Both the sensitivity and specificity of the RealAmp assay were high. The RealAmp assay could detect 10(3)CFUml(-1) within 30min. A comparative evaluation of the RealAmp assay, the API Listeria assay, and the real-time PCR assay revealed that the RealAmp assay is simpler, faster, and has a higher specificity than the other two assays. Significance and Impact of the StudyConventional culture and molecular detection methods are always time consuming and require a specific laboratory infrastructure, thereby restricting their use for the rapid detection and diagnosis of pathogens. A real-time loop-mediated isothermal amplification (RealAmp) assay performed by ESEtube scanner to rapidly detect Listeria monocytogenes isolated from food was developed. The results showed that the RealAmp assay using the tube scanner was more efficient and precise than the conventional API Listeria assay and the real-time PCR assay.
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页码：85 / 90
页数：6
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