中国兽医科学2018，Vol.48Issue(6)：700-706,7.DOI:10.16656/j.issn.1673-4696.2018.0108
禽呼肠孤病毒纳米PCR检测方法的建立与应用
Establishment and application of nano PCRassay for detection of avian reovirus
摘要
Abstract
In order to develop a rapid and sensitive method for detecting avian reovirus(ARV), a pair of primers was designed based on the conserved region of ARV S3 gene, and an ARV nano PCR assay was established. The results showed only ARV was successfully amplified with a target fragment of 332 bp in size, but the amplification results were negative for Newcastle disease virus, infectious bronchitis virus, infectious bursal disease virus, Mycoplasma gallisepticum, Marek's disease virus, infectious laryngotracheitis virus, duck plaque virus, fowl adenovirus type 4, egg drop syndrome virus, avian influenza virus(H9) and Escherichia coli. The sensitivity test showed that the sensitivity of the nano PCR was 10 times as high as that of the conventional PCR, and the lowest detection mass concentration of cDNA was 56 pg/ L. ARV nucleic acid was detected 3 times by the nano PCR, and these results were consistent. Furthermore, 30 clinical samples were detected by the nano PCR and virus isolation methods, and the coincidence rate of the two methods was 100％. The above results showed that the ARV nano PCR method was successfully established, which has high sensitivity and good specificity, and the method can be used for clinical diagnosis and epidemiological investigation of ARV infection.关键词
禽呼肠孤病毒/S3基因/纳米PCRKey words
avian reovirus/S3 gene/nano PCR分类
农业科技引用本文复制引用
郑丽,任卫科,路超,田向学,王利丽,李秀丽,鄢明华..禽呼肠孤病毒纳米PCR检测方法的建立与应用[J].中国兽医科学,2018,48(6):700-706,7.基金项目
国家重点研发计划项目(2016YFD0500800) （2016YFD0500800）
