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- Title
鉴别猪流行性腹泻病毒经典株与变异株双重纳米 RT-PCR检测方法的建立和应用.
- Authors
付琦媛; 严世杰; 王春芳; 李文傲; 马增军; 宋 涛; 芮 萍
- Abstract
In order to develop a rapid and efficient detection method to distinguish the classical and variant PEDV strains, three specific primers was designed based on S gene sequences of classical strain (CV777) and variant strain (CH-HB1-2018) in GenBank, and a duplex Nano RT-PCR method was developed based on using the gold nanoparticles as thermal conduction mesons and optimized PCR conditions. Using this method to detect classical and variant PEDV strains, PRV, PCV2, PCV3, PRRSV, and PDCoV, the results showed that the fragments from classical PEDV (747 bp) and variant PEDV (and (442 bp) were amplified,and had no cross-reaction with other five swine viruses. The diluted recombinant plasmid samples detected by this method and RT-PCR to detect the sensitivity, and the limited detection of this Nano RT-PCR was 5.68×102 copies/μL and 4.93×102 copies/μL for CV777 and CH-HB1-2018, respectively, which is 100 times higher than that of RT-PCR. The recombinant plasmid samples were detected at same and different time, which showed that the duplex Nano RT-PCR possesses the features of high repeatability and repeatability. Thirty- four samples of diarrheal piglets collected from Hebei province were detected using this method, the results showed that the coincidence rate with RT-PCR was 97.1% and the positive rate of PEDV variant strains was higher than that of classical PEDV strains. The developed duplex Nano RT- PCR can be used for the PEDV pathogen differential diagnosis and epidemiological investigation.
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2021, Vol 43, Issue 2, p145
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.202003032