title: Loop-mediated isothermal amplification assays for screening of bacterial integrons.
abstract: BACKGROUND
  The occurrence and prevalence of integrons in clinical microorganisms and their role played in antimicrobial resistance have been well studied recently. As screening and detection of integrons are concerned, current diagnostic methodologies are restricted by significant drawbacks and novel methods are required for integrons detection.
  
  
  RESULTS
  In this study, three loop-mediated isothermal amplification (LAMP) assays targeting on class 1, 2 and 3 integrons were implemented and evaluated. Optimization of these detection assays were performed, including studing on the reaction temperature, volume, time, sensitivity and specificity (both primers and targets). Application of the established LAMP assays were further verified on a total of 1082 isolates (previously identified to be 397 integron-positive and 685 integron-negative strains). According to the results, the indispensability of each primer had been confirmed and the optimal reaction temperature, volume and time were found to be 65°C, 45 min and 25 μL, respectively. As application was concerned, 361, 28 and 8 isolates carrying intI1, intI2 and intI3 yielded positive amplicons, respectively. Other 685 integron-negative bacteria were negative for the integron-screening LAMP assays, totaling the detection rate and specificity to be 100%.
  
  
  CONCLUSIONS
  The intI1-, intI2- and intI3-LAMP assays established in this study were demonstrated to be the valid and rapid detection methodologies for the screening of bacterial integrons.
authors.forename: Guangchao
authors.initials: G
authors.lastname: Yu
authors.forename: Lei
authors.initials: L
authors.lastname: Chen
authors.forename: Chii-wann
authors.initials: CW
authors.lastname: Lin
authors.forename: Bing
authors.initials: B
authors.lastname: Li
authors.forename: Hemiao
authors.initials: H
authors.lastname: Cui
authors.forename: Siyi
authors.initials: S
authors.lastname: Chen
authors.forename: Jian
authors.initials: J
authors.lastname: Miao
authors.forename: Huawei
authors.initials: H
authors.lastname: Bian
authors.forename: Dingqiang
authors.initials: D
authors.lastname: Chen
authors.forename: Yang
authors.initials: Y
authors.lastname: Deng
mesh_terms: D052160:Benzothiazoles; D015169:Colony Count, Microbial; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification*; D018076:DNA, Complementary; D003959:Diamines; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics; D005456:Fluorescent Dyes; D006358:Hot Temperature; D019426:Integrases / Q000235:genetics; D032023:Integrons; D008826:Microbial Sensitivity Tests; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods; D011804:Quinolines; D012475:Salmonella / Q000235:genetics; D012680:Sensitivity and Specificity; D012706:Serratia marcescens / Q000235:genetics; D013210:Staphylococcus / Q000235:genetics; D014734:Vibrio cholerae / Q000235:genetics
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D004269:DNA, Bacterial; D018076:DNA, Complementary; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D019426:Integrases