Accurate Detection of <i>Salmonella</i> Based on Microfluidic Chip to Avoid Aerosol Contamination
<i>Salmonella</i> is a type of common foodborne pathogen of global concern, seriously endangering human health. In molecular biological detection of <i>Salmonella</i>, the method of amplifying DNA often faces the problem of aerosol pollution. In this study, a microfluidic chi...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-12-01
|
| Series: | Foods |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2304-8158/11/23/3887 |
| _version_ | 1827643077390499840 |
|---|---|
| author | Yining Luo Shan Shan Shuanglong Wang Jinlin Li Daofeng Liu Weihua Lai |
| author_facet | Yining Luo Shan Shan Shuanglong Wang Jinlin Li Daofeng Liu Weihua Lai |
| author_sort | Yining Luo |
| collection | DOAJ |
| description | <i>Salmonella</i> is a type of common foodborne pathogen of global concern, seriously endangering human health. In molecular biological detection of <i>Salmonella</i>, the method of amplifying DNA often faces the problem of aerosol pollution. In this study, a microfluidic chip was developed to integrate loop-mediated isothermal amplification (LAMP) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system to detect <i>Salmonella</i>. The LAMP reaction solution was initially injected into the chamber to amplify at 65 °C for 20 min; the CRISPR/Cas12a reaction solution was subsequently injected to mix with the amplicons for fluorescent signal production at 43 °C for 30 min. Then, the results can be confirmed by naked eyes under 495 nm light or by a fluorescence immunochromatographic reader. The detection limit of this method for <i>Salmonella</i> DNA was 118 pg/μL. The sensitivity and specificity of this method was 100%. Furthermore, this method was used to detect <i>Salmonella</i> after enrichment for 4 h in salmon and chicken samples spiked with 30 CFU/25 g, and was verified to have a stable detection capability in real samples. The microfluidic chip integrated with the LAMP and CRISPR/Cas12a system not only provides a possibility of highly sensitive endpoint fluorescent visual detection of a foodborne pathogen, but also greatly eliminates the risk of aerosol contamination. |
| first_indexed | 2024-03-09T17:47:31Z |
| format | Article |
| id | doaj.art-7868568ce74140edadd7d20565c6f76d |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-09T17:47:31Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-7868568ce74140edadd7d20565c6f76d2023-11-24T11:00:14ZengMDPI AGFoods2304-81582022-12-011123388710.3390/foods11233887Accurate Detection of <i>Salmonella</i> Based on Microfluidic Chip to Avoid Aerosol ContaminationYining Luo0Shan Shan1Shuanglong Wang2Jinlin Li3Daofeng Liu4Weihua Lai5State Key Laboratory of Food Science and Technology, Nanchang University, 235 East Nanjing Road, Nanchang 330047, ChinaCollege of Life Science, National R&D Center for Freshwater Fish Processing, Jiangxi Normal University, Nanchang 330022, ChinaJiangxi Key Laboratory for Mass Spectrometry and Instrumentation, East China University of Technology, Nanchang 330013, ChinaCollege of Life Science, National R&D Center for Freshwater Fish Processing, Jiangxi Normal University, Nanchang 330022, ChinaJiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Province Centre for Disease Control and Prevention, 555 East Beijing Road, Nanchang 330029, ChinaState Key Laboratory of Food Science and Technology, Nanchang University, 235 East Nanjing Road, Nanchang 330047, China<i>Salmonella</i> is a type of common foodborne pathogen of global concern, seriously endangering human health. In molecular biological detection of <i>Salmonella</i>, the method of amplifying DNA often faces the problem of aerosol pollution. In this study, a microfluidic chip was developed to integrate loop-mediated isothermal amplification (LAMP) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system to detect <i>Salmonella</i>. The LAMP reaction solution was initially injected into the chamber to amplify at 65 °C for 20 min; the CRISPR/Cas12a reaction solution was subsequently injected to mix with the amplicons for fluorescent signal production at 43 °C for 30 min. Then, the results can be confirmed by naked eyes under 495 nm light or by a fluorescence immunochromatographic reader. The detection limit of this method for <i>Salmonella</i> DNA was 118 pg/μL. The sensitivity and specificity of this method was 100%. Furthermore, this method was used to detect <i>Salmonella</i> after enrichment for 4 h in salmon and chicken samples spiked with 30 CFU/25 g, and was verified to have a stable detection capability in real samples. The microfluidic chip integrated with the LAMP and CRISPR/Cas12a system not only provides a possibility of highly sensitive endpoint fluorescent visual detection of a foodborne pathogen, but also greatly eliminates the risk of aerosol contamination.https://www.mdpi.com/2304-8158/11/23/3887<i>Salmonella</i>LAMPCRISPR/Cas12avisual detection |
| spellingShingle | Yining Luo Shan Shan Shuanglong Wang Jinlin Li Daofeng Liu Weihua Lai Accurate Detection of <i>Salmonella</i> Based on Microfluidic Chip to Avoid Aerosol Contamination Foods <i>Salmonella</i> LAMP CRISPR/Cas12a visual detection |
| title | Accurate Detection of <i>Salmonella</i> Based on Microfluidic Chip to Avoid Aerosol Contamination |
| title_full | Accurate Detection of <i>Salmonella</i> Based on Microfluidic Chip to Avoid Aerosol Contamination |
| title_fullStr | Accurate Detection of <i>Salmonella</i> Based on Microfluidic Chip to Avoid Aerosol Contamination |
| title_full_unstemmed | Accurate Detection of <i>Salmonella</i> Based on Microfluidic Chip to Avoid Aerosol Contamination |
| title_short | Accurate Detection of <i>Salmonella</i> Based on Microfluidic Chip to Avoid Aerosol Contamination |
| title_sort | accurate detection of i salmonella i based on microfluidic chip to avoid aerosol contamination |
| topic | <i>Salmonella</i> LAMP CRISPR/Cas12a visual detection |
| url | https://www.mdpi.com/2304-8158/11/23/3887 |
| work_keys_str_mv | AT yiningluo accuratedetectionofisalmonellaibasedonmicrofluidicchiptoavoidaerosolcontamination AT shanshan accuratedetectionofisalmonellaibasedonmicrofluidicchiptoavoidaerosolcontamination AT shuanglongwang accuratedetectionofisalmonellaibasedonmicrofluidicchiptoavoidaerosolcontamination AT jinlinli accuratedetectionofisalmonellaibasedonmicrofluidicchiptoavoidaerosolcontamination AT daofengliu accuratedetectionofisalmonellaibasedonmicrofluidicchiptoavoidaerosolcontamination AT weihualai accuratedetectionofisalmonellaibasedonmicrofluidicchiptoavoidaerosolcontamination |