ASFV和PRV野毒及PRV疫苗毒多重纳米PCR检测方法的建立及应用
Establishment and application of multiplex nano-PCR detection method for ASFV,PRV wild virus and PRV vaccine virus
赵越 1姚来顺 2涂忠忠 3曹泽昭 1王炳量 1薛珺 1王改丽 4贺文琦 1宋德光 1兰云刚1
作者信息
- 1. 吉林大学 动物医学学院,吉林长春 130062
- 2. 吉林省疾病预防控制中心/吉林省公共卫生研究院,吉林长春 130062
- 3. 中国农业科学院长春兽医研究所,吉林长春 130122
- 4. 吉林省畜牧兽医科学研究院,吉林长春 1300621
- 折叠
摘要
为建立一种可同时检测ASFV、PRV野毒和PRV疫苗毒(缺失gE基因)的方法,本研究以ASFV的B646L基因、PRV的gE基因以及PRV的gD基因为检测对象,建立了可同时检测ASFV、PRV野毒和PRV疫苗毒的多重纳米PCR方法,并对其特异性、敏感性进行了研究.结果显示,该方法可特异性扩增ASFV、PRV野毒和PRV疫苗毒的目的基因,而检测塞内卡病毒、猪血凝性脑脊髓炎病毒、猪水疱性口炎病毒、猪圆环病毒2型时结果均呈阴性;B646L基因、gD基因和gE基因的检测限分别为4、7和8 copies/μL.此外,将商品化试剂盒检验结果与本试验建立的多重纳米PCR检测结果进行比对,结果表明符合率为100％.综上所述,本试验建立的多重纳米PCR方法具有特异性好、灵敏度高的优点,且能够同时检测ASFV、PRV野毒和PRV疫苗毒,可用于猪重大疾病的鉴别诊断和疫病防控.
Abstract
In order to establish a method for simultaneous detection of ASFV,PRV wild virus and PRV vaccine virus(gE-),a multiplex nano-PCR method for simultaneous detection of ASFV,PRV wild virus and PRV vaccine virus were established by using the B646L gene of ASFV and the gE and gD gene of PRV as the detection objects.The method's specificity and sensitivity were studied and the results showed that it could specifically amplify the target genes of ASFV,PRV wild virus and PRV vaccine strain,but the re-suits were negative when detecting Seneca virus,porcine haemagglutinating encephalomyelitis virus,porcine vesicular stomatitis virus and porcine circovirus type 2.The minimum detection limits of B646L gene,gD gene and gE gene were 4,7 and 8 copies/μL,respectively.In addition,the effectiveness of the multiplex nano-PCR detection method was compared with commonly used commercial kits for these viruses,and the results showed that the coincidence rate was 100％.In summary,the multiplex nano-PCR method established in this experiment has high specificity and sensitivity and could simultaneously detect ASFV,wild-type and vaccine PRV,which could be used for differential diagnosis and disease prevention and control of major diseases in pigs.
关键词
猪伪狂犬病病毒/非洲猪瘟病毒/多重纳米PCR检测方法/冷链猪肉Key words
pseudorabies virus/African swine fever virus/multiplex nano-PCR detection method/cold chain pork引用本文复制引用
基金项目
吉林省科技发展计划项目(0220203046SF)
出版年
2024
国家科技期刊平台
NSTL
万方数据