Simultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis
Abstract 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 describe...
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Nature Portfolio
2017
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oai:doaj.org-article:128db854a0b041bfb90b26f24c362c912021-12-02T16:06:03ZSimultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis10.1038/s41598-017-06348-z2045-2322https://doaj.org/article/128db854a0b041bfb90b26f24c362c912017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06348-zhttps://doaj.org/toc/2045-2322Abstract 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.Yixiang JiangLianhua HePingfang WuXiaolu ShiMin JiangYinghui LiYiman LinYaqun QiuFang BaiYiqun LiaoQingge LiRenLi ZhangQinghua HuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017) |
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Medicine R Science Q Yixiang Jiang Lianhua He Pingfang Wu Xiaolu Shi Min Jiang Yinghui Li Yiman Lin Yaqun Qiu Fang Bai Yiqun Liao Qingge Li RenLi Zhang Qinghua Hu Simultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis |
| description |
Abstract 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. |
| format |
article |
| author |
Yixiang Jiang Lianhua He Pingfang Wu Xiaolu Shi Min Jiang Yinghui Li Yiman Lin Yaqun Qiu Fang Bai Yiqun Liao Qingge Li RenLi Zhang Qinghua Hu |
| author_facet |
Yixiang Jiang Lianhua He Pingfang Wu Xiaolu Shi Min Jiang Yinghui Li Yiman Lin Yaqun Qiu Fang Bai Yiqun Liao Qingge Li RenLi Zhang Qinghua Hu |
| author_sort |
Yixiang Jiang |
| title |
Simultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis |
| title_short |
Simultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis |
| title_full |
Simultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis |
| title_fullStr |
Simultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis |
| title_full_unstemmed |
Simultaneous Identification of Ten Bacterial Pathogens Using the Multiplex Ligation Reaction Based on the Probe Melting Curve Analysis |
| title_sort |
simultaneous identification of ten bacterial pathogens using the multiplex ligation reaction based on the probe melting curve analysis |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/128db854a0b041bfb90b26f24c362c91 |
| work_keys_str_mv |
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| _version_ |
1718385130170482688 |