Development of multiplex PCR assay for rapid detection of Riemerella anatipestifer, Escherichia coli, and Salmonella enterica simultaneously from ducks

文献类型： 外文期刊

第一作者： Hu, Qinghai

作者： Hu, Qinghai;Tu, Jing;Han, Xiangan;Zhu, Yinyu;Ding, Chan;Yu, Shengqing;Ding, Chan;Yu, Shengqing

作者机构：

关键词： Riemerella anatipestifer;Escherichia coli;Salmonella enterica;Multiplex PCR

期刊名称：JOURNAL OF MICROBIOLOGICAL METHODS （ 影响因子：2.363； 五年影响因子：2.24 ）

ISSN： 0167-7012

年卷期： 2011 年 87 卷 1 期

页码：

收录情况： SCI

摘要： Three pathogens, Riemerella anatipestifer, Escherichia coli, and Salmonella enterica, are leading causes of bacterial fibrinous pericarditis and perihepatitis in ducks in China and worldwide. It is difficult to differentiate these pathogens when obtaining a diagnosis on clinical signs and pathological changes. The aim of this research was to develop a multiplex polymerase chain reaction (m-PCR) that could discriminate R. anatipestifer, E. colt, and S. enterica rapidly in field isolates, or detect the three bacteria in clinical samples from diseased ducks. We selected the DnaB helicase (dnaB) gene of R. anatipestifer, alkaline phosphatase (phoA) gene of E. coli and invasion protein (invA) gene of S. enterica as target genes. In optimized conditions, the limitation of detection was approximately 10(3) colony forming units (CFU) of each of these three bacterial pathogens per PCR reaction tube. The m-PCR method showed specific amplification of respective genes from R. anatipestifer, E. coli, and S. enterica. Using the m-PCR system, bacterial strains isolated from diseased ducks in our laboratory were categorized successfully, and the pathogens could also be detected in clinical samples from diseased ducks. Therefore, the m-PCR system could distinguish the three pathogens simultaneously, for identification, routine molecular diagnosis and epidemiology, in a single reaction. (C) 2011 Elsevier B.V. All rights reserved.

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