Loop-mediated isothermal amplification assays for screening of bacterial integrons

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 nov...

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Autores principales:	Yu,Guangchao, Chen,Lei, Lin,Chii-wann, Li,Bing, Cui,Hemiao, Chen,Siyi, Miao,Jian, Bian,Huawei, Chen,Dingqiang, Deng,Yang
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2014
Materias:	Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076
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spelling	oai:scielo:S0716-976020140001000762015-10-30Loop-mediated isothermal amplification assays for screening of bacterial integronsYu,GuangchaoChen,LeiLin,Chii-wannLi,BingCui,HemiaoChen,SiyiMiao,JianBian,HuaweiChen,DingqiangDeng,Yang Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron 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.info:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.47 20142014-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076en10.1186/0717-6287-47-53
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron
spellingShingle	Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron 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
description	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.
author	Yu,Guangchao Chen,Lei Lin,Chii-wann Li,Bing Cui,Hemiao Chen,Siyi Miao,Jian Bian,Huawei Chen,Dingqiang Deng,Yang
author_facet	Yu,Guangchao Chen,Lei Lin,Chii-wann Li,Bing Cui,Hemiao Chen,Siyi Miao,Jian Bian,Huawei Chen,Dingqiang Deng,Yang
author_sort	Yu,Guangchao
title	Loop-mediated isothermal amplification assays for screening of bacterial integrons
title_short	Loop-mediated isothermal amplification assays for screening of bacterial integrons
title_full	Loop-mediated isothermal amplification assays for screening of bacterial integrons
title_fullStr	Loop-mediated isothermal amplification assays for screening of bacterial integrons
title_full_unstemmed	Loop-mediated isothermal amplification assays for screening of bacterial integrons
title_sort	loop-mediated isothermal amplification assays for screening of bacterial integrons
publisher	Sociedad de Biología de Chile
publishDate	2014
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076
work_keys_str_mv	AT yuguangchao loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT chenlei loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT linchiiwann loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT libing loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT cuihemiao loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT chensiyi loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT miaojian loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT bianhuawei loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT chendingqiang loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons AT dengyang loopmediatedisothermalamplificationassaysforscreeningofbacterialintegrons
_version_	1718441528057135104

Loop-mediated isothermal amplification assays for screening of bacterial integrons

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