Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs
The global livestock industry is facing a serious threat from a widespread foot-and-mouth disease virus (FMDV) epidemic. The timely detection of FMDV can significantly mitigate its harmful effects. This study aimed to establish and evaluate a TaqMan fluorescence quantitative PCR assay to assess its...
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MDPI AG
2024-11-01
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| Series: | Veterinary Sciences |
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| Online Access: | https://www.mdpi.com/2306-7381/11/11/541 |
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| author | Changying Dong Xingyu Xiao Meiqi Wang Yajuan Sun Hui Jin Yongzhe Zhang Hongri Zhao Qianyue Cao Yanran Yang Rui Yin |
| author_facet | Changying Dong Xingyu Xiao Meiqi Wang Yajuan Sun Hui Jin Yongzhe Zhang Hongri Zhao Qianyue Cao Yanran Yang Rui Yin |
| author_sort | Changying Dong |
| collection | DOAJ |
| description | The global livestock industry is facing a serious threat from a widespread foot-and-mouth disease virus (FMDV) epidemic. The timely detection of FMDV can significantly mitigate its harmful effects. This study aimed to establish and evaluate a TaqMan fluorescence quantitative PCR assay to assess its sensitivity, specificity, reproducibility, and stability. The standard curve equation range is 6.43 × 10<sup>9</sup>–6.43 × 10<sup>1</sup> copies/µL, with an R<sup>2</sup> value of 0.996 and a standard curve equation of y = −3.586x + 36.245. The method successfully detected 64.3 copies/µL of the target gene for FMDV and exhibited high specificity for FMDV. Repeatability tests demonstrated low coefficients of variation within and between groups (<2%), indicating good reproducibility. The clinical samples analyzed using this method showed results consistent with those of the SYBR Green I RT-qPCR assay, confirming the reliability of the method. Overall, the developed test method displayed high sensitivity, specificity, reproducibility, and stability, making it suitable for the rapid diagnosis of foot-and-mouth disease in clinical settings. |
| format | Article |
| id | doaj-art-da1d50205ae94e9f83aae7a3e8a8dd96 |
| institution | Kabale University |
| issn | 2306-7381 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj-art-da1d50205ae94e9f83aae7a3e8a8dd962024-11-26T18:24:52ZengMDPI AGVeterinary Sciences2306-73812024-11-01111154110.3390/vetsci11110541Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in PigsChangying Dong0Xingyu Xiao1Meiqi Wang2Yajuan Sun3Hui Jin4Yongzhe Zhang5Hongri Zhao6Qianyue Cao7Yanran Yang8Rui Yin9College of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaCollege of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin 132101, ChinaThe global livestock industry is facing a serious threat from a widespread foot-and-mouth disease virus (FMDV) epidemic. The timely detection of FMDV can significantly mitigate its harmful effects. This study aimed to establish and evaluate a TaqMan fluorescence quantitative PCR assay to assess its sensitivity, specificity, reproducibility, and stability. The standard curve equation range is 6.43 × 10<sup>9</sup>–6.43 × 10<sup>1</sup> copies/µL, with an R<sup>2</sup> value of 0.996 and a standard curve equation of y = −3.586x + 36.245. The method successfully detected 64.3 copies/µL of the target gene for FMDV and exhibited high specificity for FMDV. Repeatability tests demonstrated low coefficients of variation within and between groups (<2%), indicating good reproducibility. The clinical samples analyzed using this method showed results consistent with those of the SYBR Green I RT-qPCR assay, confirming the reliability of the method. Overall, the developed test method displayed high sensitivity, specificity, reproducibility, and stability, making it suitable for the rapid diagnosis of foot-and-mouth disease in clinical settings.https://www.mdpi.com/2306-7381/11/11/541foot-and-mouth disease virusTaqMan real-time RT-qPCRsystem optimizationdetection methods |
| spellingShingle | Changying Dong Xingyu Xiao Meiqi Wang Yajuan Sun Hui Jin Yongzhe Zhang Hongri Zhao Qianyue Cao Yanran Yang Rui Yin Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs Veterinary Sciences foot-and-mouth disease virus TaqMan real-time RT-qPCR system optimization detection methods |
| title | Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs |
| title_full | Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs |
| title_fullStr | Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs |
| title_full_unstemmed | Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs |
| title_short | Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs |
| title_sort | development and application of a taqman rt qpcr for the detection of foot and mouth disease virus in pigs |
| topic | foot-and-mouth disease virus TaqMan real-time RT-qPCR system optimization detection methods |
| url | https://www.mdpi.com/2306-7381/11/11/541 |
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