摘要
为了对禽腺病毒4型(FAdV-4)引发的鸡心包积水-肝炎综合征的病原学监测和疫情诊断提供技术支撑,本研究经对部分发表在NCBI的禽腺病毒4型序列进行比对,在其六邻体基因上找到一段相对保守序列,并设计出PCR引物和TaqMan探针。以FAdV-4 HB1510株作为标准品,通过反应条件优化,建立了FAdV-4荧光定量PCR检测方法,并对其敏感性、特异性和重复性进行了分析。结果显示,该方法的灵敏度为10 EID_(50)/mL,为常规PCR方法的10倍;组内和组间重复性良好;与鸡减蛋综合征病毒、大肠杆菌等禽类病原的核酸无交叉反应。通过对65份临床样品进行检测,FAdV-4荧光定量PCR和常规PCR的检测符合率为90.7%。因此,FAdV-4荧光定量PCR的快速、敏感、特异等优点使其在禽腺病毒4型的早期检测及预防、控制等方面有较好的应用前景。
In order to provide technical support for pathogenic monitoring and epidemic diagnosis of hydropericardium hepatitis syndrome caused by avian adenovirus type 4(FAdV-4),genome sequences of FAdV-4 were downloaded from NCBI and a relatively conserved sequence in its hexon gene was identified after comparison,from which PCR primers and TaqMan probes were designed.The sensitivity,specificity,and repeatability of the FAdV-4 fluorescent quantitative PCR were assessed using FAdV-4 HB1510 strain as a template.The results showed that the FAdV-4 fluorescent quantitative PCR had a sensitivity of 10 EID_(50)/mL,which was 10-fold higher than that of conventional PCR,displayed good reproducibility,and demonstrated no cross-reaction with nucleic acids of other avian pathogens,such as egg drop syndrome virus and Escherichia coli.Based on examination of 65 clinical samples,FAdV-4 fluorescent quantitative PCR and conventional PCR had 90.7%agreement.Therefore,the FAdV-4 fluorescent quantitative PCR provides a sensitive,specific and rapid tool for early detection and improved monitoring of avian adenovirus type 4.
作者
王国康
卢琴
贾妙妙
李丽
冯贺龙
邵华斌
罗青平
曾驰
温国元
WANG Guo-kang;LU Qin;JIA Miao-miao;LI Li;FENG He-long;SHAO Hua-bin;LUO Qing-ping;ZENG Chi;WEN Guo-yuan(School of Biology and Pharmaceutical Engineering,Wuhan Polytechnic University,Wuhan 430023,China;Institute of Animal Husbandry and Veterinary,Hubei Academy of Agricultural Sciences,Wuhan 430064,China;Key Laboratory of Prevention and Control Agents for Animal Bacteriosis of Ministry of Agriculture and Rural Affairs,Wuhan 430064,China;Hubei Provincial Key Laboratory of Livestock and Poultry Pathogen Microbiology,Wuhan 430064,China)
出处
《中国兽医杂志》
CAS
北大核心
2022年第2期22-26,共5页
Chinese Journal of Veterinary Medicine
基金
国家重点研发计划(2017YFD0501104)
国家现代农业产业技术体系建设专项资金(CARS-43-G13)
国家自然科学基金项目(31873018,31670157)
湖北省自然科学基金杰出青年人才项目(2019CFA071)
湖北省对外科技合作项目(2019AHB066)。
