Development and application of real-time polymerase chain reaction to detect Vibrio cholerae O1 and O139 in river water / 中华流行病学杂志
Chinese Journal of Epidemiology
; (12): 768-771, 2007.
Article
en Zh
| WPRIM
| ID: wpr-294240
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To develop a real-time SYBR Green polymerase chain reaction (PCR) for detection of Vibrio cholerae serogroups O1 and O139, and to evaluate its reliability through detection of estuary water samples.</p><p><b>METHODS</b>O antigen rfb genes specific for O1 and O139 were used for the design of PCR primers. The real-time SYBR Green PCR system in detecting O1 and O139 specific rfb genes in one tube was developed, and its sensitivity, specificity and reproducibility were evaluated. The ability of the real-time PCR in detection of estuary water samples was compared with the routine PCR and bacteria isolation.</p><p><b>RESULTS</b>The amplification of O1 or O139 specific target gene could be detected according to the melt curve temperature of amplicons. No amplification was observed in the templates of other 10 non-cholerae vibrios. When comparing to the real-time PCR to bacteria isolation in detection of 524 estuary water samples, it showed high sensitivity, plus also positive in real-time PCR detection among all the samples in which bacteria of O1 or O139 were isolated.</p><p><b>CONCLUSION</b>The real-time SYBR Green PCR could be used as the first step of rapid environment screen of V. cholerae in water samples thus might enhance the efficiency of isolation in screening of large amount of water samples.</p>
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Monitoreo del Ambiente
/
Reacción en Cadena de la Polimerasa
/
Reproducibilidad de los Resultados
/
Sensibilidad y Especificidad
/
Vibrio cholerae O1
/
Vibrio cholerae O139
/
Ríos
/
Genes Bacterianos
/
Métodos
/
Microbiología
Tipo de estudio:
Diagnostic_studies
Idioma:
Zh
Revista:
Chinese Journal of Epidemiology
Año:
2007
Tipo del documento:
Article