Developing a novel TaqMan qPCR assay for optimizing Salmonella Pullorum detection in chickens
Salmonella Pullorum, the causative agent of pullorum disease, posing a significant threat to the global production of poultry meat and eggs. However, existing detection methods have substantial limitations in efficiency and accuracy. Herein, we developed a genomic deletion-targeted TaqMan qPCR assay...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2025-12-01
|
| Series: | Veterinary Quarterly |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/01652176.2025.2454473 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832580031443894272 |
|---|---|
| author | Hao Wang Xiamei Kang Longhai Yu Haijian Wang Anja Müller Corinna Kehrenberg Yan Li Min Yue |
| author_facet | Hao Wang Xiamei Kang Longhai Yu Haijian Wang Anja Müller Corinna Kehrenberg Yan Li Min Yue |
| author_sort | Hao Wang |
| collection | DOAJ |
| description | Salmonella Pullorum, the causative agent of pullorum disease, posing a significant threat to the global production of poultry meat and eggs. However, existing detection methods have substantial limitations in efficiency and accuracy. Herein, we developed a genomic deletion-targeted TaqMan qPCR assay for identification of Salmonella Pullorum, enabling precise differentiation from other Salmonella serovars. The assay’s detection limit was 5 copies/μL of plasmid and 4 CFU/μL of bacterial DNA. Furthermore, we collected 676 chicken samples from an established infection model to compare the performance of the TaqMan qPCR assay with traditional bacterial culturing and antibody-based detection approaches. With superior sensitivity and specificity, the newly developed method detected over 80% of positive chickens, significantly outperforming the two conventional methods. Moreover, we proposed a combined framework that incorporates the advantages of TaqMan qPCR assay and antibody detection method, further enhancing the detection rate of positives to 92%. Additionally, to address the frequent aerosol contamination of amplification products in laboratory settings, we devised an easy-to-deploy anti-contamination system based on T7 exonuclease. Overall, the T7 exonuclease-assisted TaqMan qPCR assay will not only upgrade the current detection for pullorum disease, but also exemplify the feasibility of targeting specific genomic deletions for pathogen detection. |
| format | Article |
| id | doaj-art-9ddc72adfb954c91839ace6f718f0cd5 |
| institution | Kabale University |
| issn | 0165-2176 1875-5941 |
| language | English |
| publishDate | 2025-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Veterinary Quarterly |
| spelling | doaj-art-9ddc72adfb954c91839ace6f718f0cd52025-01-30T11:18:19ZengTaylor & Francis GroupVeterinary Quarterly0165-21761875-59412025-12-0145111310.1080/01652176.2025.2454473Developing a novel TaqMan qPCR assay for optimizing Salmonella Pullorum detection in chickensHao Wang0Xiamei Kang1Longhai Yu2Haijian Wang3Anja Müller4Corinna Kehrenberg5Yan Li6Min Yue7Key Laboratory of Systems Health Science of Zhejiang Province, School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, ChinaDepartment of Veterinary Medicine, Zhejiang University College of Animal Sciences, Hangzhou, ChinaDepartment of Veterinary Medicine, Zhejiang University College of Animal Sciences, Hangzhou, ChinaHainan Institute of Zhejiang University, Sanya, ChinaInstitute for Veterinary Food Science, Faculty of Veterinary Medicine, Justus Liebig University Giessen, Giessen, GermanyInstitute for Veterinary Food Science, Faculty of Veterinary Medicine, Justus Liebig University Giessen, Giessen, GermanyDepartment of Veterinary Medicine, Zhejiang University College of Animal Sciences, Hangzhou, ChinaKey Laboratory of Systems Health Science of Zhejiang Province, School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, ChinaSalmonella Pullorum, the causative agent of pullorum disease, posing a significant threat to the global production of poultry meat and eggs. However, existing detection methods have substantial limitations in efficiency and accuracy. Herein, we developed a genomic deletion-targeted TaqMan qPCR assay for identification of Salmonella Pullorum, enabling precise differentiation from other Salmonella serovars. The assay’s detection limit was 5 copies/μL of plasmid and 4 CFU/μL of bacterial DNA. Furthermore, we collected 676 chicken samples from an established infection model to compare the performance of the TaqMan qPCR assay with traditional bacterial culturing and antibody-based detection approaches. With superior sensitivity and specificity, the newly developed method detected over 80% of positive chickens, significantly outperforming the two conventional methods. Moreover, we proposed a combined framework that incorporates the advantages of TaqMan qPCR assay and antibody detection method, further enhancing the detection rate of positives to 92%. Additionally, to address the frequent aerosol contamination of amplification products in laboratory settings, we devised an easy-to-deploy anti-contamination system based on T7 exonuclease. Overall, the T7 exonuclease-assisted TaqMan qPCR assay will not only upgrade the current detection for pullorum disease, but also exemplify the feasibility of targeting specific genomic deletions for pathogen detection.https://www.tandfonline.com/doi/10.1080/01652176.2025.2454473Salmonella Pullorumpullorum diseasechickenTaqMan qPCRgenomic deletionmolecular detection |
| spellingShingle | Hao Wang Xiamei Kang Longhai Yu Haijian Wang Anja Müller Corinna Kehrenberg Yan Li Min Yue Developing a novel TaqMan qPCR assay for optimizing Salmonella Pullorum detection in chickens Veterinary Quarterly Salmonella Pullorum pullorum disease chicken TaqMan qPCR genomic deletion molecular detection |
| title | Developing a novel TaqMan qPCR assay for optimizing Salmonella Pullorum detection in chickens |
| title_full | Developing a novel TaqMan qPCR assay for optimizing Salmonella Pullorum detection in chickens |
| title_fullStr | Developing a novel TaqMan qPCR assay for optimizing Salmonella Pullorum detection in chickens |
| title_full_unstemmed | Developing a novel TaqMan qPCR assay for optimizing Salmonella Pullorum detection in chickens |
| title_short | Developing a novel TaqMan qPCR assay for optimizing Salmonella Pullorum detection in chickens |
| title_sort | developing a novel taqman qpcr assay for optimizing salmonella pullorum detection in chickens |
| topic | Salmonella Pullorum pullorum disease chicken TaqMan qPCR genomic deletion molecular detection |
| url | https://www.tandfonline.com/doi/10.1080/01652176.2025.2454473 |
| work_keys_str_mv | AT haowang developinganoveltaqmanqpcrassayforoptimizingsalmonellapullorumdetectioninchickens AT xiameikang developinganoveltaqmanqpcrassayforoptimizingsalmonellapullorumdetectioninchickens AT longhaiyu developinganoveltaqmanqpcrassayforoptimizingsalmonellapullorumdetectioninchickens AT haijianwang developinganoveltaqmanqpcrassayforoptimizingsalmonellapullorumdetectioninchickens AT anjamuller developinganoveltaqmanqpcrassayforoptimizingsalmonellapullorumdetectioninchickens AT corinnakehrenberg developinganoveltaqmanqpcrassayforoptimizingsalmonellapullorumdetectioninchickens AT yanli developinganoveltaqmanqpcrassayforoptimizingsalmonellapullorumdetectioninchickens AT minyue developinganoveltaqmanqpcrassayforoptimizingsalmonellapullorumdetectioninchickens |