title: Detection on integron carried gene cassettes from pathogens by loop mediated isothermal amplification assays.
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.
authors.affiliation: Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: yuguangchao2017@163.com.
authors.forename: Guangchao
authors.initials: G
authors.lastname: Yu
authors.affiliation: Department of Dermatology and Venerology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, PR China. Electronic address: yecongxiu1603@163.com.
authors.forename: Congxiu
authors.initials: C
authors.lastname: Ye
authors.affiliation: Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: fuqiang1421@163.com.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Fu
authors.affiliation: Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: liujuzhen8806@163.com.
authors.forename: Juzhen
authors.initials: J
authors.lastname: Liu
authors.affiliation: Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: fanxiaoyi6203@163.com.
authors.forename: Xiaoyi
authors.initials: X
authors.lastname: Fan
authors.affiliation: Clinical Laboratory Center, The First Affiliated Hospital of Jinan University, Guangzhou 510630, PR China. Electronic address: huangyunzu@163.com.
authors.forename: Yunzu
authors.initials: Y
authors.lastname: Huang
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, 382 Zhonghuan Road East, Guangzhou 510006, PR China. Electronic address: zhoushishui2017@163.com.
authors.forename: Shishui
authors.initials: S
authors.lastname: Zhou
mesh_terms: D001419:Bacteria / Q000235:genetics*; D017931:DNA Primers; D004269:DNA, Bacterial; D005798:Genes, Bacterial / Q000235:genetics*; D006358:Hot Temperature; D032023:Integrons / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial
keywords: Bacterial integrons; Gene cassettes characterization; Loop-mediated isothermal amplification (LAMP)