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Establishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmealestablishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmeal
Daojin Yu
Fujian Agriculture and Forestry University
Corresponding Author
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This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at AMB Express.
Salmonella spp. is a high-risk bacterial pathogen that is monitored in imported animal-derived feedstuffs. Serratiafonticola is the bacterial species most frequently confused with Salmonella spp. in traditional identification methods based on biochemical characteristics, which are time-consuming and labor-intensive, and thus unsuitable for daily inspection and quarantine work. In this study, we established a duplex real-time qPCR method with invA- and gyrB-specific primers and probes corresponding to Salmonella spp. and S. fonticola. The method could simultaneously detect both pathogens in imported feedstuffs, with a minimum limit of detection for Salmonella spp. and S. fonticola of 197 copies/SL and 145 copies/SL, respectively (correlation coefficient R2 = 0.999 in both cases). The amplification efficiency for Salmonella spp. and S. fonticola was 98.346% and 96.49%, respectively. Detection of clinical samples was consistent with method GB/T 13091-2018, and all seven artificially contaminated imported feed samples were positively identified. Thus, the developed duplex real-time qPCR assay displays high specificity and sensitivity, and can be used for the rapid and accurate detection of genomic DNA from Salmonella spp. and S. fonticola within hours. This represents a significant improvement in the efficiency of detection of both pathogens in imported feedstuffs.
Keywords
Salmonella spp., S. fonticola, Duplex Real-time qPCR, Feed safety
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CURRENT STATUS: Published
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Posted 21 Sep, 2020
Editorial decision: Minor Revision
On 24 Oct, 2020
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Received 23 Oct, 2020
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On 17 Oct, 2020
Reviewers invited
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On 05 Oct, 2020
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On 04 Oct, 2020
Editor invited
On 04 Oct, 2020
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On 24 Nov, 2020
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This preprint is under consideration at AMB Express. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Original article
Establishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmealestablishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmeal
Daojin Yu
Fujian Agriculture and Forestry University
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Version (no preprint)
Posted 21 Sep, 2020
Editorial decision: Minor Revision
On 24 Oct, 2020
Review #1 received
Received 23 Oct, 2020
Reviewer #1 agreed
On 17 Oct, 2020
Reviewers invited
Invitations sent on 08 Oct, 2020
Editor assigned
On 05 Oct, 2020
Submission checks complete
On 04 Oct, 2020
Editor invited
On 04 Oct, 2020
This version was not preprinted
Version 1
Posted 21 Sep, 2020
Published
On 24 Nov, 2020
No community comments so far
Editor assigned
On 28 Sep, 2020
Editor invited
On 27 Sep, 2020
Submission checks complete
On 16 Sep, 2020
First submitted
On 14 Sep, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at AMB Express.
Salmonella spp. is a high-risk bacterial pathogen that is monitored in imported animal-derived feedstuffs. Serratiafonticola is the bacterial species most frequently confused with Salmonella spp. in traditional identification methods based on biochemical characteristics, which are time-consuming and labor-intensive, and thus unsuitable for daily inspection and quarantine work. In this study, we established a duplex real-time qPCR method with invA- and gyrB-specific primers and probes corresponding to Salmonella spp. and S. fonticola. The method could simultaneously detect both pathogens in imported feedstuffs, with a minimum limit of detection for Salmonella spp. and S. fonticola of 197 copies/SL and 145 copies/SL, respectively (correlation coefficient R2 = 0.999 in both cases). The amplification efficiency for Salmonella spp. and S. fonticola was 98.346% and 96.49%, respectively. Detection of clinical samples was consistent with method GB/T 13091-2018, and all seven artificially contaminated imported feed samples were positively identified. Thus, the developed duplex real-time qPCR assay displays high specificity and sensitivity, and can be used for the rapid and accurate detection of genomic DNA from Salmonella spp. and S. fonticola within hours. This represents a significant improvement in the efficiency of detection of both pathogens in imported feedstuffs.
Figure 1
Figure 2
Figure 3
Figure 4