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- Title
基于重组酶介导等温扩增技术的羊口疮病毒核酸检测方法 的建立及初步应用.
- Authors
张学勇; 简莹娜; 李志; 郭志宏; 严永盛; 朵红
- Abstract
In order to establish a rapid detection method for Orf virus (ORFV) nucleic acid, this study designed specific primers based on the conserved sequence of ORFV B2L gene, and preliminarily established a recombinase-mediated isothermal amplification technology by optimizing the reaction temperature and time (RAA) ORFV detection method. The optimization test results show that the method has the best detection effect at 37 ℃ for 40 min. This method was used to detect viral nucleic acids with similar clinical symptoms, such as ORFV, goat pox virus, O-type and A-type foot-and-mouth disease virus, Peste des Petits Ruminants virus, and goat parainfluenza virus type 3. In addition to being positive, the detection results of other viruses in sheep were all negative, with strong specificity. The plasmid standard was diluted 10 times (1×109 copies/μL to 1×100 copies/μL) as a template, and the RAA method established in this study was used for detection. The results showed that this method had the lowest detection rate for ORFV plasmid standards. The limit is 1×104 copies/μL, and the sensitivity is high. Using the method established in this study to detect 95 clinical samples (15 tissue samples and 80 blood samples), the results showed that the RAA method can quickly detect clinical samples, and the positive rates of tissue samples and blood samples were 33.33% ( 5/15) and 6.25% (5/80), the coincidence rate of the detection results with the common PCR detection method is 100%, indicating that the RAA method established in this experiment can meet the detection requirements of clinical samples. The ORFV RAA method established in this study has the characteristics of simple operation, strong specificity, and rapid response, which provides a new technical choice for the rapid detection of ORFV.
- Subjects
PARAINFLUENZA viruses; COINCIDENCE circuits; PESTE des petits ruminants; NUCLEIC acids; POXVIRUSES; BLOOD sampling; FOOT &; mouth disease; PLANT viruses
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2023, Vol 45, Issue 3, p271
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.202206013