■番鸭细小病毒(MDPV),鹅细小病毒(GPV),鸭圆环病毒,(DuCV)和鸭腺病毒3(DAdV-3)是引起鸭高发病率和死亡率的重要病原体,给养鸭业造成巨大的经济损失。
■本研究,一种四重一步实时定量PCR方法检测MDPV,GPV,DuCV,并且开发了DAdV-3。
■结果表明,该测定与其他家禽病原体没有交叉反应性[鸭瘟病毒(DPV),鸭热病毒(DTMUV),H6禽流感病毒(H6AIV),新鸭呼肠孤病毒(NDRV),新城疫病毒(NDV),H4禽流感病毒(H4AIV),大肠杆菌(E.大肠杆菌),番鸭呼肠孤病毒(MDRV),产卵综合征病毒(EDSV),多杀巴斯德氏菌(P.multocida)].灵敏度结果表明,MDPV的检测限,GPV,DuCV,和DAdV-3是10,10,1和10拷贝/μl,分别;方法内和方法间的变异系数为1-2%;线性范围(109至103拷贝/µL)表明MDPV的R2值,GPV,DuCV,和DAdV-3分别为0.9975、0.998、0.9964和0.996。四重实时定量PCR方法的效率为90.30%,101.10%,90.72%,MDPV为90.57%,GPV,DuCV,和DAdV-3。采用建立的四重实时定量PCR方法和已报道的检测方法,同时检测了2022年6月至2023年7月在部分鸭香肠中收集的396份临床标本。MDPV的检出率,GPV,DuCV,DAdV-3为8.33%(33/396),17.93%(71/396),33.58%(133/396),和29.04%(115/396),分别。这些测定之间的一致性大于99.56%。
■开发的四重实时定量PCR测定法可以准确检测感染鸭子的这四种病毒,提供一个快速的,敏感,临床检测的具体和准确的技术。
UNASSIGNED: Muscovy duck parvovirus (MDPV), Goose parvovirus (GPV), Duck circovirus, (DuCV) and Duck adenovirus 3 (DAdV-3) are important pathogens that cause high morbidity and mortality in ducks, causing huge economic loss for the duck industry.
UNASSIGNED: The present study, a
quadruplex one-step real time quantitative PCR method for the detection of MDPV, GPV, DuCV, and DAdV-3 was developed.
UNASSIGNED: The results showed that assay had no cross-reactivity with other poultry pathogens [Duck plague virus (DPV), Duck tembusu virus (DTMUV), H6 avian influenza virus (H6 AIV), New duck reovirus (NDRV), Newcastle disease virus (NDV), H4 avian influenza virus (H4 AIV), Escherichia coli (E. coli), Muscovy duck reovirus (MDRV), Egg drop syndrome virus (EDSV), Pasteurella multocida (P. multocida)]. The sensitivity result showed that the limits of detection for MDPV, GPV, DuCV, and DAdV-3 were 10, 10, 1 and 10 copies/µl, respectively; The coefficients of variation intra- and inter-method was 1-2%; The range of linear (109 to 103 copies/µL) demonstrated the R2 values for MDPV, GPV, DuCV, and DAdV-3 as 0.9975, 0.998, 0.9964, and 0.996, respectively. The
quadruplex real time quantitative PCR method efficiency was 90.30%, 101.10%, 90.72%, and 90.57% for MDPV, GPV, DuCV, and DAdV-3, respectively. 396 clinical specimens collected in some duck sausages from June 2022 to July 2023 were simultaneously detected using the established
quadruplex real time quantitative PCR method and the reported assays. The detection rates for MDPV, GPV, DuCV, and DAdV-3 were 8.33% (33/396), 17.93% (71/396), 33.58% (133/396), and 29.04% (115/396), respectively. The agreement between these assays was greater than 99.56%.
UNASSIGNED: The developed
quadruplex real-time quantitative PCR assay can accurately detect these four viruses infecting ducks, providing a rapid, sensitive, specific and accurate technique for clinical testing.