A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of <i>Shigella</i> and <i>Bacillus cereus</i> in Food
<i>Shigella</i> and <i>Bacillus cereus</i> are two common foodborne pathogens that cause intestinal diseases and seriously affect human life and health. Traditional microbiological culture methods are time-consuming and laborious, and polymerase chain reaction (PCR)-based met...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-05-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/12/9/1889 |
| _version_ | 1797602644131315712 |
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| author | Shuna Xiang Hanyue Zhang Xiaoyan Cha Yingting Lin Ying Shang |
| author_facet | Shuna Xiang Hanyue Zhang Xiaoyan Cha Yingting Lin Ying Shang |
| author_sort | Shuna Xiang |
| collection | DOAJ |
| description | <i>Shigella</i> and <i>Bacillus cereus</i> are two common foodborne pathogens that cause intestinal diseases and seriously affect human life and health. Traditional microbiological culture methods are time-consuming and laborious, and polymerase chain reaction (PCR)-based methods rely on expensive thermal cyclers and lengthy reaction times. In this study, on the basis of the specific gene <i>ipaH7</i> of <i>Shigella</i> and the virulence gene <i>nheABC</i> of <i>B. cereus</i>, a duplex detection system was established for the first time by using the recombinase polymerase amplification technique (D-RPA). After optimization, D-RPA could be effectively amplified at 42 °C for 25 min with excellent specificity, and the detection limits of D-RPA for <i>Shigella</i> and <i>B. cereus</i> in artificially contaminated samples were 2.7 × 10<sup>1</sup> and 5.2 × 10<sup>2</sup> CFU/mL, respectively. This study provides a certain research basis for multiple detection with RPA, an isothermal amplification technology. Furthermore, it lays a good foundation for high-throughput rapid detection of foodborne pathogens. |
| first_indexed | 2024-03-11T04:19:31Z |
| format | Article |
| id | doaj.art-bd56a8b963224d508082f11b89f17ddc |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-11T04:19:31Z |
| publishDate | 2023-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-bd56a8b963224d508082f11b89f17ddc2023-11-17T22:56:14ZengMDPI AGFoods2304-81582023-05-01129188910.3390/foods12091889A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of <i>Shigella</i> and <i>Bacillus cereus</i> in FoodShuna Xiang0Hanyue Zhang1Xiaoyan Cha2Yingting Lin3Ying Shang4Faculty of Food Science and Engineering, Kunming University of Science and Technology, Yunnan 650500, ChinaFaculty of Food Science and Engineering, Kunming University of Science and Technology, Yunnan 650500, ChinaFaculty of Food Science and Engineering, Kunming University of Science and Technology, Yunnan 650500, ChinaFaculty of Food Science and Engineering, Kunming University of Science and Technology, Yunnan 650500, ChinaFaculty of Food Science and Engineering, Kunming University of Science and Technology, Yunnan 650500, China<i>Shigella</i> and <i>Bacillus cereus</i> are two common foodborne pathogens that cause intestinal diseases and seriously affect human life and health. Traditional microbiological culture methods are time-consuming and laborious, and polymerase chain reaction (PCR)-based methods rely on expensive thermal cyclers and lengthy reaction times. In this study, on the basis of the specific gene <i>ipaH7</i> of <i>Shigella</i> and the virulence gene <i>nheABC</i> of <i>B. cereus</i>, a duplex detection system was established for the first time by using the recombinase polymerase amplification technique (D-RPA). After optimization, D-RPA could be effectively amplified at 42 °C for 25 min with excellent specificity, and the detection limits of D-RPA for <i>Shigella</i> and <i>B. cereus</i> in artificially contaminated samples were 2.7 × 10<sup>1</sup> and 5.2 × 10<sup>2</sup> CFU/mL, respectively. This study provides a certain research basis for multiple detection with RPA, an isothermal amplification technology. Furthermore, it lays a good foundation for high-throughput rapid detection of foodborne pathogens.https://www.mdpi.com/2304-8158/12/9/1889recombinase polymerase amplificationduplex detection<i>Shigella</i><i>Bacillus cereus</i> |
| spellingShingle | Shuna Xiang Hanyue Zhang Xiaoyan Cha Yingting Lin Ying Shang A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of <i>Shigella</i> and <i>Bacillus cereus</i> in Food Foods recombinase polymerase amplification duplex detection <i>Shigella</i> <i>Bacillus cereus</i> |
| title | A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of <i>Shigella</i> and <i>Bacillus cereus</i> in Food |
| title_full | A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of <i>Shigella</i> and <i>Bacillus cereus</i> in Food |
| title_fullStr | A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of <i>Shigella</i> and <i>Bacillus cereus</i> in Food |
| title_full_unstemmed | A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of <i>Shigella</i> and <i>Bacillus cereus</i> in Food |
| title_short | A New Duplex Recombinase Polymerase Amplification (D-RPA) Method for the Simultaneous and Rapid Detection of <i>Shigella</i> and <i>Bacillus cereus</i> in Food |
| title_sort | new duplex recombinase polymerase amplification d rpa method for the simultaneous and rapid detection of i shigella i and i bacillus cereus i in food |
| topic | recombinase polymerase amplification duplex detection <i>Shigella</i> <i>Bacillus cereus</i> |
| url | https://www.mdpi.com/2304-8158/12/9/1889 |
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