||Yu, Guangchao; Chen, Lei; Lin, Chii-wann; Li, Bing; Cui, Hemiao; Chen, Siyi; Miao, Jian; Bian, Huawei; Chen, Dingqiang; Deng, Yang.|
||Loop-mediated isothermal amplification assays for screening of bacterial integrons|
||Biol. Res;47:1-10, 2014. ilus, tab.
||National 973-Plan of China; . International Science & Technology Cooperation Program; . Science and Technology Planning Project of Guangdong Province, China; . National Natural Science Foundation of China; . National Science and Technology Support Program; . National Outstanding Doctoral Dissertation Funding; . Guangdong Outstanding Doctoral Dissertation Funding; . China Postdoctoral Science Foundation funded; . Fundamental Research Funds for the Central Universities; . Fund for Outstanding Youth of Anhui Academy of Agricultural Sciences.
||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.|
||DNA Bacteriano/isolamento & purificação|
Técnicas de Amplificação de Ácido Nucleico/métodos
Contagem de Colônia Microbiana
Testes de Sensibilidade Microbiana
Reação em Cadeia da Polimerase/métodos
Sensibilidade e Especificidade
Primers do DNA
Eletroforese em Gel de Ágar
| Tipo de Publ:
||Research Support, Non-U.S. Gov't|
||CL1.1 - Biblioteca Central|