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Simultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis.
Jiang, Yixiang; He, Lianhua; Wu, Pingfang; Shi, Xiaolu; Jiang, Min; Li, Yinghui; Lin, Yiman; Qiu, Yaqun; Bai, Fang; Liao, Yiqun; Li, Qingge; Zhang, RenLi; Hu, Qinghua.
Afiliación
  • Jiang Y; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • He L; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • Wu P; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • Shi X; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • Jiang M; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • Li Y; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • Lin Y; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • Qiu Y; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • Bai F; School of Life Sciences, Shenzhen University, Shenzhen, 518000, Guangdong, China.
  • Liao Y; School of Life Sciences, Xiamen University, Xiamen, 361005, Fujian Province, China.
  • Li Q; School of Life Sciences, Xiamen University, Xiamen, 361005, Fujian Province, China.
  • Zhang R; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China.
  • Hu Q; Shenzhen Center for Disease Control and Prevention, Shenzhen, 518020, China. huqinghua03@163.com.
Sci Rep ; 7(1): 5902, 2017 07 19.
Article en En | MEDLINE | ID: mdl-28724932
Pathogenic Vibrio spp., Aeromonas spp. and Plesiomonas shigelloides are associated with human gastroenteritis and wound infections, as well as fish diseases. The comprehensive and accurate identification of these pathogens is crucial for the current public health. The present study describes the development of a multiplex assay for the simultaneous identification of ten bacterial pathogens in a single reaction by using a multiplex ligation reaction based on probe melting curve analysis (MLMA). The specificity for target genes was 100%, as assessed with a panel of 67 bacterial pathogens, which indicated no cross-reactions. The detection limit of this assay ranged from 0.8 × 107 CFU/mL to 1.5 × 108 CFU/mL at the pure bacterial culture level and from 0.1 ng to 1.0 ng at the DNA level. The MLMA assay was used to detect ten species of pathogens in 269 clinical and seafood samples, and for further validation, the results were compared with the conventional culture method. The results indicated greater than 90% sensitivity and 100% specificity for each bacterial pathogen tested, and the kappa correlation for all the pathogens ranged from 0.95 to 1.00. Overall, this assay is well suited for public health laboratories for its high throughput, accuracy, and low cost.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Bacterias / Sondas de ADN / Reacción en Cadena de la Polimerasa Multiplex / Desnaturalización de Ácido Nucleico Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Sci Rep Año: 2017 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Bacterias / Sondas de ADN / Reacción en Cadena de la Polimerasa Multiplex / Desnaturalización de Ácido Nucleico Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Sci Rep Año: 2017 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido