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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Autor*in:	Guangchao Yu [verfasserIn] Lei Chen [verfasserIn] Chii-wann Lin [verfasserIn] Bing Li [verfasserIn] Hemiao Cui [verfasserIn] Siyi Chen [verfasserIn] Jian Miao [verfasserIn] Huawei Bian [verfasserIn] Dingqiang Chen [verfasserIn] Yang Deng [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2014

Schlagwörter:	Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron

Übergeordnetes Werk:	In: Biological Research - BMC, 2018, 47(2014), 0, Seite 10
Übergeordnetes Werk:	volume:47 ; year:2014 ; number:0 ; pages:10

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1186/0717-6287-47-53

Katalog-ID:	DOAJ043364403

Internformat


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520			\|a 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.
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allfields	10.1186/0717-6287-47-53 doi (DE-627)DOAJ043364403 (DE-599)DOAJ33c60460b33b43d7ad818c083e36920c DE-627 ger DE-627 rakwb eng QH301-705.5 Guangchao Yu verfasserin aut Loop-mediated isothermal amplification assays for screening of bacterial integrons 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron Biology (General) Lei Chen verfasserin aut Chii-wann Lin verfasserin aut Bing Li verfasserin aut Hemiao Cui verfasserin aut Siyi Chen verfasserin aut Jian Miao verfasserin aut Huawei Bian verfasserin aut Dingqiang Chen verfasserin aut Yang Deng verfasserin aut In Biological Research BMC, 2018 47(2014), 0, Seite 10 (DE-627)329975374 (DE-600)2048380-6 07176287 nnns volume:47 year:2014 number:0 pages:10 https://doi.org/10.1186/0717-6287-47-53 kostenfrei https://doaj.org/article/33c60460b33b43d7ad818c083e36920c kostenfrei http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076&lng=en&tlng=en kostenfrei https://doaj.org/toc/0716-9760 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 47 2014 0 10
spelling	10.1186/0717-6287-47-53 doi (DE-627)DOAJ043364403 (DE-599)DOAJ33c60460b33b43d7ad818c083e36920c DE-627 ger DE-627 rakwb eng QH301-705.5 Guangchao Yu verfasserin aut Loop-mediated isothermal amplification assays for screening of bacterial integrons 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron Biology (General) Lei Chen verfasserin aut Chii-wann Lin verfasserin aut Bing Li verfasserin aut Hemiao Cui verfasserin aut Siyi Chen verfasserin aut Jian Miao verfasserin aut Huawei Bian verfasserin aut Dingqiang Chen verfasserin aut Yang Deng verfasserin aut In Biological Research BMC, 2018 47(2014), 0, Seite 10 (DE-627)329975374 (DE-600)2048380-6 07176287 nnns volume:47 year:2014 number:0 pages:10 https://doi.org/10.1186/0717-6287-47-53 kostenfrei https://doaj.org/article/33c60460b33b43d7ad818c083e36920c kostenfrei http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076&lng=en&tlng=en kostenfrei https://doaj.org/toc/0716-9760 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 47 2014 0 10
allfields_unstemmed	10.1186/0717-6287-47-53 doi (DE-627)DOAJ043364403 (DE-599)DOAJ33c60460b33b43d7ad818c083e36920c DE-627 ger DE-627 rakwb eng QH301-705.5 Guangchao Yu verfasserin aut Loop-mediated isothermal amplification assays for screening of bacterial integrons 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron Biology (General) Lei Chen verfasserin aut Chii-wann Lin verfasserin aut Bing Li verfasserin aut Hemiao Cui verfasserin aut Siyi Chen verfasserin aut Jian Miao verfasserin aut Huawei Bian verfasserin aut Dingqiang Chen verfasserin aut Yang Deng verfasserin aut In Biological Research BMC, 2018 47(2014), 0, Seite 10 (DE-627)329975374 (DE-600)2048380-6 07176287 nnns volume:47 year:2014 number:0 pages:10 https://doi.org/10.1186/0717-6287-47-53 kostenfrei https://doaj.org/article/33c60460b33b43d7ad818c083e36920c kostenfrei http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076&lng=en&tlng=en kostenfrei https://doaj.org/toc/0716-9760 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 47 2014 0 10
allfieldsGer	10.1186/0717-6287-47-53 doi (DE-627)DOAJ043364403 (DE-599)DOAJ33c60460b33b43d7ad818c083e36920c DE-627 ger DE-627 rakwb eng QH301-705.5 Guangchao Yu verfasserin aut Loop-mediated isothermal amplification assays for screening of bacterial integrons 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron Biology (General) Lei Chen verfasserin aut Chii-wann Lin verfasserin aut Bing Li verfasserin aut Hemiao Cui verfasserin aut Siyi Chen verfasserin aut Jian Miao verfasserin aut Huawei Bian verfasserin aut Dingqiang Chen verfasserin aut Yang Deng verfasserin aut In Biological Research BMC, 2018 47(2014), 0, Seite 10 (DE-627)329975374 (DE-600)2048380-6 07176287 nnns volume:47 year:2014 number:0 pages:10 https://doi.org/10.1186/0717-6287-47-53 kostenfrei https://doaj.org/article/33c60460b33b43d7ad818c083e36920c kostenfrei http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076&lng=en&tlng=en kostenfrei https://doaj.org/toc/0716-9760 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 47 2014 0 10
allfieldsSound	10.1186/0717-6287-47-53 doi (DE-627)DOAJ043364403 (DE-599)DOAJ33c60460b33b43d7ad818c083e36920c DE-627 ger DE-627 rakwb eng QH301-705.5 Guangchao Yu verfasserin aut Loop-mediated isothermal amplification assays for screening of bacterial integrons 2014 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier 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. Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron Biology (General) Lei Chen verfasserin aut Chii-wann Lin verfasserin aut Bing Li verfasserin aut Hemiao Cui verfasserin aut Siyi Chen verfasserin aut Jian Miao verfasserin aut Huawei Bian verfasserin aut Dingqiang Chen verfasserin aut Yang Deng verfasserin aut In Biological Research BMC, 2018 47(2014), 0, Seite 10 (DE-627)329975374 (DE-600)2048380-6 07176287 nnns volume:47 year:2014 number:0 pages:10 https://doi.org/10.1186/0717-6287-47-53 kostenfrei https://doaj.org/article/33c60460b33b43d7ad818c083e36920c kostenfrei http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076&lng=en&tlng=en kostenfrei https://doaj.org/toc/0716-9760 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 47 2014 0 10
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callnumber-first	Q - Science
author	Guangchao Yu
spellingShingle	Guangchao Yu misc QH301-705.5 misc Loop-mediated isothermal amplification (LAMP) misc Integron screening misc Bacterial integrons misc Class 1 integron misc Class 2 integron misc Class 3 integron misc Biology (General) Loop-mediated isothermal amplification assays for screening of bacterial integrons
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topic_title	QH301-705.5 Loop-mediated isothermal amplification assays for screening of bacterial integrons Loop-mediated isothermal amplification (LAMP) Integron screening Bacterial integrons Class 1 integron Class 2 integron Class 3 integron
topic	misc QH301-705.5 misc Loop-mediated isothermal amplification (LAMP) misc Integron screening misc Bacterial integrons misc Class 1 integron misc Class 2 integron misc Class 3 integron misc Biology (General)
topic_unstemmed	misc QH301-705.5 misc Loop-mediated isothermal amplification (LAMP) misc Integron screening misc Bacterial integrons misc Class 1 integron misc Class 2 integron misc Class 3 integron misc Biology (General)
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title	Loop-mediated isothermal amplification assays for screening of bacterial integrons
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title_full	Loop-mediated isothermal amplification assays for screening of bacterial integrons
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title_sort	loop-mediated isothermal amplification assays for screening of bacterial integrons
callnumber	QH301-705.5
title_auth	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.
abstractGer	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.
abstract_unstemmed	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.
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title_short	Loop-mediated isothermal amplification assays for screening of bacterial integrons
url	https://doi.org/10.1186/0717-6287-47-53 https://doaj.org/article/33c60460b33b43d7ad818c083e36920c http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100076&lng=en&tlng=en https://doaj.org/toc/0716-9760
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author2	Lei Chen Chii-wann Lin Bing Li Hemiao Cui Siyi Chen Jian Miao Huawei Bian Dingqiang Chen Yang Deng
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up_date	2024-07-03T17:09:23.479Z
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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%. 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Loop-mediated isothermal amplification assays for screening of bacterial integrons

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