Chousalkar K K, Cheetham B F, Roberts J R
Animal Science, School of Environmental and Rural Science, University of New England, Armidale, New South Wales 2351, Australia.
J Virol Methods. 2009 Jan;155(1):67-71. doi: 10.1016/j.jviromet.2008.09.028. Epub 2008 Nov 14.
In the present study, LNA-probe based real-time PCR was designed for the detection and absolute quantification of infectious bronchitis virus (IBV) from the oviduct of unvaccinated and vaccinated hens after IBV challenge. Using a recombinant plasmid standard, the detection limit of the reaction was found to be 10 copies and independent assay runs showed reproducible Ct values. Amongst the unvaccinated hens, the virus could be detected between 6 and 20 days post-infection (p.i.), with a peak of viral load between 10 and 14 days p.i. The virus was also detectable in the oviduct of vaccinated, challenged hens although the viral load was much lower compared to the viral load in the oviduct of unvaccinated, challenged hens. This indicates that rearing phase vaccination can offer significant protection of the fully functional oviduct against a pathogenic strain of IBV. The present test will be useful for the rapid identification of IBV directly from clinical samples. Most vaccination trials investigating the efficacy of vaccines for layer and breeder hens have been conducted based on the respiratory tract response. Evaluation of viral load from the oviduct of vaccinated and unvaccinated hens is an efficient method for assessing oviduct protection in commercial laying hens.
在本研究中，设计了基于锁核酸（LNA）探针的实时荧光定量聚合酶链反应（PCR），用于检测和绝对定量接种传染性支气管炎病毒（IBV）疫苗和未接种疫苗的母鸡在受到IBV攻击后输卵管中的病毒。使用重组质粒标准品，发现该反应的检测限为10个拷贝，并且独立的检测运行显示出可重复的Ct值。在未接种疫苗的母鸡中，感染后6至20天可检测到病毒，病毒载量在感染后10至14天达到峰值。在接种疫苗并受到攻击的母鸡的输卵管中也可检测到病毒，尽管与未接种疫苗且受到攻击的母鸡输卵管中的病毒载量相比要低得多。这表明育雏期接种疫苗可以为功能完全正常的输卵管提供显著保护，使其免受致病性IBV毒株的侵害。本试验将有助于直接从临床样本中快速鉴定IBV。大多数研究蛋鸡和种母鸡疫苗效力的接种试验都是基于呼吸道反应进行的。评估接种疫苗和未接种疫苗的母鸡输卵管中的病毒载量是评估商业蛋鸡输卵管保护情况的有效方法。
