Detection on integron carried gene cassettes from pathogens by loop mediated isothermal amplification assays. (June 2017)

• Record Type: Journal Article
• Authors: Yu, Guangchao; Ye, Congxiu; Fu, Qiang; Liu, Juzhen; Fan, Xiaoyi; Huang, Yunzu; Zhou, Shishui
• Is Part Of: Microbial pathogenesis. Volume 107(2017)
• Page Start: 304
• Page End: 308
• Publication Date: 2017-06
• Subjects: Loop-mediated isothermal amplification (LAMP) -- Bacterial integrons -- Gene cassettes characterization; Pathogenic microorganisms -- Periodicals; Pathology, Molecular -- Periodicals; Communicable Diseases -- microbiology -- Periodicals; Communicable Diseases -- parasitology -- Periodicals; Micro-organismes pathogènes -- Périodiques; Pathologie moléculaire -- Périodiques; Electronic journals; 616.9041
• DOI: 10.1016/j.micpath.2017.04.006
• Languages: English
• ISSNs: 0882-4010

Abstract

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 bla VIM2 . 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. Highlights: Six valid and rapid LAMP assays targeting on integron genes were developed. Gene dfrA12, orfF, aadA2, dfrA17, aadA5 and bla VIM2 were used as target genes. 272 positive and 685 negative isolates were applied on LAMP with 100% detection rate and specificity.