Development of a novel SYBR green I-based quantitative RT-PCR assay for Senecavirus A detection in clinical samples of pigs
| Autor: | Xiaobo Wen, Huichen Guo, Jin Xu, Suyu Mu, Hu Dong, Sahibzada Waheed Abdullah, Shiqi Sun, Yun Zhang, Mei Li, Zhidong Teng, Shichong Han |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Swine
Picornaviridae Biology Diamines 03 medical and health sciences chemistry.chemical_compound Vesicular Stomatitis Swine Vesicular Disease Limit of Detection Animals Benzothiazoles Molecular Biology Senecavirus A Swine vesicular disease 030304 developmental biology Gel electrophoresis Swine Diseases 0303 health sciences Picornaviridae Infections 030306 microbiology Reverse Transcriptase Polymerase Chain Reaction Cell Biology Virology Reverse transcription polymerase chain reaction Real-time polymerase chain reaction chemistry Nucleic acid SYBR Green I Quinolines Capsid Proteins |
| Zdroj: | Molecular and cellular probes. 53 |
| ISSN: | 1096-1194 |
| Popis: | Porcine vesicular disease caused by Senecavirus A (SVA) is a newly emerging disease in many countries. Based on clinical signs only, it is very challenging to distinguish SVA infection from other similar diseases, such as foot and mouth disease, swine vesicular disease, and vesicular stomatitis. Therefore, it is crucial to establish a detection assay for the clinical diagnosis of SVA infection. In this study, a pair of specific primers were designed based on the highly conserved L/VP4 gene sequence of SVA. The established SYBR green I-based quantitative reverse transcription polymerase chain reaction (qRT-PCR) method was used to detect SVA nucleic acids in clinical samples. The limit of detection SVA nucleic acids by qRT-PCR was 6.4 × 101 copies/μL, which was significantly more sensitive than that by gel electrophoresis of 6.4 × 103 copes/μL. This assay was specific and had no cross-reaction with other seven swine viruses. Using SYBR green I-based qRT-PCR, the SVA positive rates in experimental animal samples and field samples were 67.60% (96/142) and 80% (24/30) respectively. The results demonstrate that SYBR green I-based qRT-PCR is a rapid and specific method for the clinical diagnosis and epidemiological investigation of related vesicular diseases caused by SVA. |
| Databáze: | OpenAIRE |
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