Development of a TaqMan-based real-time RT-PCR assay for the detection of SADS-CoV associated with severe diarrhea disease in pigs
| العنوان: | Development of a TaqMan-based real-time RT-PCR assay for the detection of SADS-CoV associated with severe diarrhea disease in pigs |
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| المؤلفون: | Jingyun Ma, Xiangbin Zhang, Yang Bai, Gui-hua Chen, Jiao-ling Wu, Jian Cheng, Di Li, Zixian Wu, Kai-jie Mai, Zhihai Zhou, Ruiting Wu, Yuan Sun, Ling Zhou |
| المصدر: | Journal of Virological Methods |
| بيانات النشر: | Elsevier B.V., 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, Diarrhea, Swine, viruses, 030106 microbiology, Disease, Biology, medicine.disease_cause, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Virus, Article, 03 medical and health sciences, Virology, Quantification, Diagnosis, TaqMan, medicine, Animals, Severe diarrhea, Coronavirus, Detection limit, Swine Diseases, Alphacoronavirus, Reproducibility of Results, 030104 developmental biology, Real-time polymerase chain reaction, Specific primers, TaqMan-based real-time RT-PCR, SADS-CoV, Coronavirus Infections |
| الوصف: | Highlights • We established a TaqMan-based real-time RT-PCR assay for the detection of a novel swine diarrhea virus SADS-CoV. • The assay in this study was of high sensitivity, specificity and reproducibility. • We compared the accuracy between conventional PCR and TaqMan-based RT-PCR assay with adequate clinical samples. • This approach will help to provide knowledge on epidemiology and pathogenesis studies of SADS-CoV. Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a novel coronavirus which was first reported in southern China in 2017. It can cause severe diarrhea disease in pigs. In order to detect this new emerging virus rapidly and reliably, a TaqMan-based real-time RT-PCR assay was established in this study. Specific primers and probe were designed and synthesized based on the conserved region within the N gene of the viral genome. Results showed that the lowest limit of detection was 3.0 × 101 copies/μL. This approach was specific for SADS-CoV, and there were no cross-reaction observed against other 15 swine viruses. It was 10 times more sensitive than the conventional PCR and gave higher SADS-CoV positive detection rate (70.69%, 123/174) than the conventional PCR (51.15%, 89/174) from clinical samples. These data indicated that the TaqMan-based real-time RT-PCR assay established here was an effective method with high sensitivity, specificity and reproducibility for faster and more accurate detection and quantification of SADS-CoV. |
| اللغة: | English |
| تدمد: | 1879-0984 0166-0934 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b4a9d0246bf6fee5c54df021d5af9758 http://europepmc.org/articles/PMC7113665 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....b4a9d0246bf6fee5c54df021d5af9758 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 18790984 01660934 |
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