SciELO - Scientific Electronic Library Online

 
vol.47Human testis-expressed sequence 101 is limitedly distributed in germinal epithelium of testis and disappears in seminoma índice de autoresíndice de materiabúsqueda de artículos
Home Pagelista alfabética de revistas  

Servicios Personalizados

Revista

Articulo

Indicadores

Links relacionados

  • En proceso de indezaciónCitado por Google
  • No hay articulos similaresSimilares en SciELO
  • En proceso de indezaciónSimilares en Google

Compartir


Biological Research

versión impresa ISSN 0716-9760

Resumen

YU, Guangchao et al. Loop-mediated isothermal amplification assays for screening of bacterial integrons. Biol. Res. [online]. 2014, vol.47, pp.1-10. ISSN 0716-9760.  http://dx.doi.org/10.1186/0717-6287-47-53.

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.

Palabras clave : Loop-mediated isothermal amplification (LAMP); Integron screening; Bacterial integrons; Class 1 integron; Class 2 integron; Class 3 integron.

        · texto en Inglés     · Inglés ( pdf )