Loop-mediated isothermal amplification assays for screening of bacterial integrons
Biol. Res
;
47: 1-10, 2014. ilus, tab
Artículo
en Inglés
| LILACS
| ID: biblio-950749
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.
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
ADN Bacteriano
/
Técnicas de Amplificación de Ácido Nucleico
/
Integrones
Tipo de estudio:
Estudio diagnóstico
/
Factores de riesgo
/
Estudio de tamizaje
Idioma:
Inglés
Revista:
Biol. Res
Asunto de la revista:
Biologia
Año:
2014
Tipo del documento:
Artículo
País de afiliación:
China
/
Taiwán
Institución/País de afiliación:
Anhui Academy of Agricultural Science/CN
/
First Affiliated Hospital of Guangzhou Medical College/CN
/
Jinan University/CN
/
National Taiwan University/TW
/
South China University of Technology/CN
/
Yat-sen University/CN
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