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Microb Pathog. 2017 Jun;107:304-308. doi: 10.1016/j.micpath.2017.04.006. Epub 2017 Apr 6.

Detection on integron carried gene cassettes from pathogens by loop mediated isothermal amplification assays.

Author information

1
Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: yuguangchao2017@163.com.
2
Department of Dermatology and Venerology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, PR China. Electronic address: yecongxiu1603@163.com.
3
Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: fuqiang1421@163.com.
4
Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: liujuzhen8806@163.com.
5
Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: fanxiaoyi6203@163.com.
6
Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: huangyunzu@163.com.
7
School of Bioscience and Bioengineering, South China University of Technology, 382 Zhonghuan Road East, Guangzhou 510006, PR China. Electronic address: zhoushishui2017@163.com.

Abstract

In this study, a number of frequently detected gene cassettes from bacterial integrons have been detected and characterized by rapid and simple loop-mediated isothermal amplification (LAMP) assays. Six gene cassettes commonly found in class 1 integrons were studied, including dfrA12, dfrA17, aadA2, aadA5, orfF, and blaVIM2. Primers design, sensitivity, specificity, optimization of each LAMP assay, as well as application of the developed 6 individual LAMP assays on a large scale of bacteria, had been conducted. The optimal amplification was obtained with temperature as 65 °C, reaction time span as 45 min and volume as 25 μl. For application, 272 isolates with various gene cassettes yielded expectable positive amplicons and other 685 integron-negative bacteria showed negative results for the LAMP assays, totaling 100% detection rate and specificity.

KEYWORDS:

Bacterial integrons; Gene cassettes characterization; Loop-mediated isothermal amplification (LAMP)

PMID:
28392412
DOI:
10.1016/j.micpath.2017.04.006
[Indexed for MEDLINE]

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