Development of a multiplex PCR assay for the simultaneous and rapid detection of six pathogenic bacteria in poultry

文献类型： 外文期刊

第一作者： Wang, Zhihao

作者： Wang, Zhihao;Zuo, Jiakun;Hu, Jiangang;Jiang, Wei;Mi, Rongsheng;Huang, Yan;Chen, Zhaoguo;Han, Xiangan;Wang, Zhihao;Hu, Jiangang;Qi, Kezong;Gong, Jiansen;Phouthapane, Vanhnaseng;Wang, Chen

作者机构：

关键词： Avian; Six pathogenic bacteria; Multiplex PCR assay; Specificity

期刊名称：AMB EXPRESS （ 影响因子：3.298； 五年影响因子：3.427 ）

ISSN： 2191-0855

年卷期： 2019 年 9 卷 1 期

页码：

收录情况： SCI

摘要： Escherichia coli, Pasteurella multocida, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella spp. and Staphylococcus aureus are six bacterial pathogens of avian. However, these pathogens may cause many similar pathological changes, resulting in clinical isolates that are difficult to quickly and simultaneously detect and identify. Here, a multiplex polymerase chain reaction (m-PCR) assay is reported to rapidly identify targets genes (phoA, KMT1, ureR, toxA, invA, and nuc) of these six pathogens in clinical samples. Six pairs of specific primers were designed. The optimal reaction conditions, specificity, and sensitivity of the m-PCR assay were investigated. The results showed that betaine remarkably improved amplification of the target genes. Specific test results showed that all six pathogens were detected by the proposed m-PCR protocol without cross-amplification with viruses or parasites. Sensitivity test results showed that the m-PCR system could amplify the six target genes from bacterial genomes or cultures with template amounts of 500 pg or 2.8-8.6 x 10(3) colony forming units, respectively. Furthermore, the six bacterial pathogens isolated from the infected tissue samples were successfully identified. The proposed m-PCR assay is a useful tool to monitor and diagnose bacterial infection in birds with high specificity, sensitivity and throughput.

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