Establishment of a TaqMan Quantitative Real-Time PCR for Detecting <i>Lawsonia intracellularis</i>
Porcine proliferative enteropathy (PPE) is an infectious disease in pigs, caused by <i>Lawsonia intracellularis</i> (LI), affecting their intestines during growth and finishing stages, leading to higher production costs. Current detection methods for LI face two main challenges, delayed...
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MDPI AG
2025-05-01
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| Series: | Veterinary Sciences |
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| Online Access: | https://www.mdpi.com/2306-7381/12/5/450 |
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| author | Zhiqiang Hu Ranran Lai Wei Xu Ran Guan Zhimin Zhang Guangwen Yan Guiying Hao |
| author_facet | Zhiqiang Hu Ranran Lai Wei Xu Ran Guan Zhimin Zhang Guangwen Yan Guiying Hao |
| author_sort | Zhiqiang Hu |
| collection | DOAJ |
| description | Porcine proliferative enteropathy (PPE) is an infectious disease in pigs, caused by <i>Lawsonia intracellularis</i> (LI), affecting their intestines during growth and finishing stages, leading to higher production costs. Current detection methods for LI face two main challenges, delayed results and high costs, making them impractical for large-scale pig farming epidemiological surveys. This study developed a TaqMan-qPCR method using specific probes and primers based on the LI aspartate ammonia lyase genes from GenBank, completing detection in just 45 min. After optimizing reaction conditions, sensitivity analysis revealed that the detection limit of this method was 4.6 copies/μL targeting standard plasmids. The results of the specificity analysis showed no cross-reactivity with other common porcine pathogens, highlighting its specificity. The inter- and intra-group coefficients of variation were both <1%, indicating high reproducibility. Furthermore, the TaqMan-qPCR demonstrated 100% relative sensitivity, and a 92.50% compliance rate compared to conventional PCR, suggesting it could be a complement to the conventional PCR method. In summary, the TaqMan-qPCR method established in this study is not only suitable for epidemiological investigations and early qualitative and quantitative diagnosis of proliferative enteropathy in pigs, but it is also valuable for studying the biological characteristics of LI. |
| format | Article |
| id | doaj-art-ffecc6b281a3432eb719fa8191d47e2d |
| institution | OA Journals |
| issn | 2306-7381 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj-art-ffecc6b281a3432eb719fa8191d47e2d2025-08-20T02:33:51ZengMDPI AGVeterinary Sciences2306-73812025-05-0112545010.3390/vetsci12050450Establishment of a TaqMan Quantitative Real-Time PCR for Detecting <i>Lawsonia intracellularis</i>Zhiqiang Hu0Ranran Lai1Wei Xu2Ran Guan3Zhimin Zhang4Guangwen Yan5Guiying Hao6Key Laboratory of Animal Epidemic Disease Detection and Prevention in Panxi District, College of Animal Science, Xichang University, Xichang 615013, ChinaShandong New Hope Liuhe Agriculture and Animal Husbandry Technology Co., Ltd., Dezhou 253034, ChinaMOA Key Laboratory of Animal Virology, Center for Veterinary Sciences, Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, ChinaKey Laboratory of Animal Epidemic Disease Detection and Prevention in Panxi District, College of Animal Science, Xichang University, Xichang 615013, ChinaKey Laboratory of Animal Epidemic Disease Detection and Prevention in Panxi District, College of Animal Science, Xichang University, Xichang 615013, ChinaKey Laboratory of Animal Epidemic Disease Detection and Prevention in Panxi District, College of Animal Science, Xichang University, Xichang 615013, ChinaKey Laboratory of Animal Epidemic Disease Detection and Prevention in Panxi District, College of Animal Science, Xichang University, Xichang 615013, ChinaPorcine proliferative enteropathy (PPE) is an infectious disease in pigs, caused by <i>Lawsonia intracellularis</i> (LI), affecting their intestines during growth and finishing stages, leading to higher production costs. Current detection methods for LI face two main challenges, delayed results and high costs, making them impractical for large-scale pig farming epidemiological surveys. This study developed a TaqMan-qPCR method using specific probes and primers based on the LI aspartate ammonia lyase genes from GenBank, completing detection in just 45 min. After optimizing reaction conditions, sensitivity analysis revealed that the detection limit of this method was 4.6 copies/μL targeting standard plasmids. The results of the specificity analysis showed no cross-reactivity with other common porcine pathogens, highlighting its specificity. The inter- and intra-group coefficients of variation were both <1%, indicating high reproducibility. Furthermore, the TaqMan-qPCR demonstrated 100% relative sensitivity, and a 92.50% compliance rate compared to conventional PCR, suggesting it could be a complement to the conventional PCR method. In summary, the TaqMan-qPCR method established in this study is not only suitable for epidemiological investigations and early qualitative and quantitative diagnosis of proliferative enteropathy in pigs, but it is also valuable for studying the biological characteristics of LI.https://www.mdpi.com/2306-7381/12/5/450porcine proliferative enteropathy<i>Lawsonia intracellularis</i>TaqMan-qPCRclinical detection |
| spellingShingle | Zhiqiang Hu Ranran Lai Wei Xu Ran Guan Zhimin Zhang Guangwen Yan Guiying Hao Establishment of a TaqMan Quantitative Real-Time PCR for Detecting <i>Lawsonia intracellularis</i> Veterinary Sciences porcine proliferative enteropathy <i>Lawsonia intracellularis</i> TaqMan-qPCR clinical detection |
| title | Establishment of a TaqMan Quantitative Real-Time PCR for Detecting <i>Lawsonia intracellularis</i> |
| title_full | Establishment of a TaqMan Quantitative Real-Time PCR for Detecting <i>Lawsonia intracellularis</i> |
| title_fullStr | Establishment of a TaqMan Quantitative Real-Time PCR for Detecting <i>Lawsonia intracellularis</i> |
| title_full_unstemmed | Establishment of a TaqMan Quantitative Real-Time PCR for Detecting <i>Lawsonia intracellularis</i> |
| title_short | Establishment of a TaqMan Quantitative Real-Time PCR for Detecting <i>Lawsonia intracellularis</i> |
| title_sort | establishment of a taqman quantitative real time pcr for detecting i lawsonia intracellularis i |
| topic | porcine proliferative enteropathy <i>Lawsonia intracellularis</i> TaqMan-qPCR clinical detection |
| url | https://www.mdpi.com/2306-7381/12/5/450 |
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