中国动物传染病学报2026，Vol.34Issue(2)：70-78,9.
鸭甲肝病毒3型、鸭星状病毒3型、鸭腺病毒3型多重PCR检测方法的建立及初步应用
Development and Preliminary Application of a Multiplex PCR for Detection of DHAV-3,DAstV-3 and DAdV-3
摘要
Abstract
In order to develop a multiplex PCR method for simultaneous detection of Duck hepatitis A virus-3(DHAV-3),Duck astrovirus-3(DAstV-3)and Duck adenovirus-3(DAdV-3),three sets of specific primers were designed based on the conserved regions of their genomes.The resulting amplified products were 298,505 and 668 bp,respectively.The multiplex PCR assay was developed and optimized for testing DHAV-3,DAstV-3 and DAdV-3.The specificity,sensitivity and reproducibility of the method were then validated for clinical application.The results showed that the multiplex PCR method developed here simultaneously amplified the target fragments of DHAV-3,DAstV-3 and DAdV-3 and there were no amplification products of other common duck pathogens.The detection limits of the three viruses were 1.0×103,1.0×104 and 1.0×104 copies/μL,respectively.Additionally,the method showed excellent repeatability with coefficients of three tests using the same templates.The multiplex PCR method was applied to test the clinical samples and the results showed 100%agreement with routine single(RT-)PCR tests.In conclusion,the multiplex PCR method developed here was specific,sensitive and reproducible and could be used for clinical detection and epidemiological investigation of DHAV-3,DAstV-3 and DAdV-3.关键词
鸭甲肝病毒3型/鸭星状病毒3型/鸭腺病毒3型/多重PCRKey words
Duck hepatitis A virus-3/Duck astrovirus-3/Duck adenovirus-3/multiplex PCR分类
农业科技引用本文复制引用
钟敏,程龙飞,刘荣昌,万春和,傅光华,黄瑜..鸭甲肝病毒3型、鸭星状病毒3型、鸭腺病毒3型多重PCR检测方法的建立及初步应用[J].中国动物传染病学报,2026,34(2):70-78,9.基金项目
福建省禽病防治重点实验室开放基金(FKADL-2024-02) （FKADL-2024-02）
福建省农业科学院自由探索科技创新项目(ZYTS2024014) （ZYTS2024014）
