Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood
Foodborne pathogens can cause foodborne illness. In reality, one food sample may carry more than one pathogen. A rapid, sensitive, and multiple target method for bacteria detection is crucial in food safety. For the simultaneous detection of <i>Staphylococcus aureus</i>, <i>Vibrio...
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MDPI AG
2020-03-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/9/3/278 |
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| author | Biao Ma Jiali Li Kai Chen Xiaoping Yu Chuanxin Sun Mingzhou Zhang |
| author_facet | Biao Ma Jiali Li Kai Chen Xiaoping Yu Chuanxin Sun Mingzhou Zhang |
| author_sort | Biao Ma |
| collection | DOAJ |
| description | Foodborne pathogens can cause foodborne illness. In reality, one food sample may carry more than one pathogen. A rapid, sensitive, and multiple target method for bacteria detection is crucial in food safety. For the simultaneous detection of <i>Staphylococcus aureus</i>, <i>Vibrio parahaemolyticus</i>, and <i>Salmonella</i> Enteritidis, multi-objective recombinase polymerase amplification (RPA) combined with a lateral flow dipstick (LFD) was developed in this study. The whole process, including amplification and reading, can be completed in 15 min at 37 °C. The detection limits were 2.6 × 10<sup>1</sup> CFU/mL for <i>Staphylococcus aureus</i>, 7.6 × 10<sup>1</sup> CFU/mL for <i>Vibrio parahaemolyticus</i>, and 1.29 × 10<sup>1</sup> CFU/mL for <i>Salmonella</i> Enteritidis. Moreover, colored signal intensities on test lines were measured by a test strip reader to achieve quantitative detection for <i>Staphylococcus aureus</i> (R<sup>2</sup> = 0.9903), <i>Vibrio parahaemolyticus</i> (R<sup>2</sup> = 0.9928), and <i>Salmonella</i> Enteritidis (R<sup>2</sup> = 0.9945). In addition, the method demonstrated good recoveries (92.00%−107.95%) in the testing of spiked food samples. Therefore, the multiplex LFD-RPA assay is a feasible method for the rapid, sensitive, and quantitative detection of bacterial pathogens in seafood. |
| first_indexed | 2024-12-12T05:59:32Z |
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| language | English |
| last_indexed | 2024-12-12T05:59:32Z |
| publishDate | 2020-03-01 |
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| record_format | Article |
| series | Foods |
| spelling | doaj.art-3a3e6269f02346ac802c9b7392c471922022-12-22T00:35:28ZengMDPI AGFoods2304-81582020-03-019327810.3390/foods9030278foods9030278Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in SeafoodBiao Ma0Jiali Li1Kai Chen2Xiaoping Yu3Chuanxin Sun4Mingzhou Zhang5Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, ChinaZhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, ChinaZhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, ChinaZhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, ChinaDepartment of Plant Biology, Uppsala BioCenter, Linnean Centre for Plant Biology, Swedish University of Agricultural Science (SLU), SE-75007 Uppsala, SwedenZhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, ChinaFoodborne pathogens can cause foodborne illness. In reality, one food sample may carry more than one pathogen. A rapid, sensitive, and multiple target method for bacteria detection is crucial in food safety. For the simultaneous detection of <i>Staphylococcus aureus</i>, <i>Vibrio parahaemolyticus</i>, and <i>Salmonella</i> Enteritidis, multi-objective recombinase polymerase amplification (RPA) combined with a lateral flow dipstick (LFD) was developed in this study. The whole process, including amplification and reading, can be completed in 15 min at 37 °C. The detection limits were 2.6 × 10<sup>1</sup> CFU/mL for <i>Staphylococcus aureus</i>, 7.6 × 10<sup>1</sup> CFU/mL for <i>Vibrio parahaemolyticus</i>, and 1.29 × 10<sup>1</sup> CFU/mL for <i>Salmonella</i> Enteritidis. Moreover, colored signal intensities on test lines were measured by a test strip reader to achieve quantitative detection for <i>Staphylococcus aureus</i> (R<sup>2</sup> = 0.9903), <i>Vibrio parahaemolyticus</i> (R<sup>2</sup> = 0.9928), and <i>Salmonella</i> Enteritidis (R<sup>2</sup> = 0.9945). In addition, the method demonstrated good recoveries (92.00%−107.95%) in the testing of spiked food samples. Therefore, the multiplex LFD-RPA assay is a feasible method for the rapid, sensitive, and quantitative detection of bacterial pathogens in seafood.https://www.mdpi.com/2304-8158/9/3/278recombinase polymerase amplificationlateral flow dipstickmultiplex detectionquantitative detection<i>staphylococcus aureus</i><i>vibrio parahaemolyticus</i><i>salmonella</i> enteritidis |
| spellingShingle | Biao Ma Jiali Li Kai Chen Xiaoping Yu Chuanxin Sun Mingzhou Zhang Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood Foods recombinase polymerase amplification lateral flow dipstick multiplex detection quantitative detection <i>staphylococcus aureus</i> <i>vibrio parahaemolyticus</i> <i>salmonella</i> enteritidis |
| title | Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood |
| title_full | Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood |
| title_fullStr | Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood |
| title_full_unstemmed | Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood |
| title_short | Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood |
| title_sort | multiplex recombinase polymerase amplification assay for the simultaneous detection of three foodborne pathogens in seafood |
| topic | recombinase polymerase amplification lateral flow dipstick multiplex detection quantitative detection <i>staphylococcus aureus</i> <i>vibrio parahaemolyticus</i> <i>salmonella</i> enteritidis |
| url | https://www.mdpi.com/2304-8158/9/3/278 |
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