Freick Markus, Müller Hermann, Raue Rüdiger
Institute for Virology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 29, D-04103 Leipzig, Germany.
J Virol Methods. 2008 Mar;148(1-2):226-31. doi: 10.1016/j.jviromet.2007.11.003. Epub 2008 Jan 22.
Infections of young racing pigeons with pigeon herpesvirus (PiHV), fowl adenovirus (FAdV) and pigeon circovirus (PiCV) are reported frequently. The role of these viruses in the pathogenesis of a disease complex called young pigeon disease syndrome (YPDS) is generally accepted. All of these viruses cause inclusion bodies in the liver so liver samples are particularly useful for the detection of infection. Consequently a multiplex polymerase chain reaction (PCR) was developed for the detection of PiHV, FAdV and PiCV in liver samples from racing pigeons. The detection limits were 10(1) genome equivalents for the detection of PiHV and PiCV and 10(3) genome equivalents for FAdV. The absence of PCR inhibitors was shown by the detection of cytochrome B gene as an internal control. No PCR products were amplified from related herpes and circoviruses or negative controls, demonstrating the specificity of the multiplex PCR. The addition of cellular DNA from liver samples or Q-solution to the reaction mix had no influence on its sensitivity. The usefulness of the multiplex PCR was demonstrated by re-investigation of liver samples from young racing pigeons previously tested positive by uniplex PCRs.
赛鸽幼鸽感染鸽疱疹病毒（PiHV）、禽腺病毒（FAdV）和鸽圆环病毒（PiCV）的情况屡有报道。这些病毒在一种名为幼鸽病综合征（YPDS）的疾病复合体发病机制中的作用已得到普遍认可。所有这些病毒都会在肝脏中形成包涵体，因此肝脏样本对于检测感染尤为有用。因此，开发了一种多重聚合酶链反应（PCR），用于检测赛鸽肝脏样本中的PiHV、FAdV和PiCV。PiHV和PiCV的检测限为10¹基因组当量，FAdV的检测限为10³基因组当量。通过检测细胞色素B基因作为内部对照，表明不存在PCR抑制剂。未从相关疱疹病毒和圆环病毒或阴性对照中扩增出PCR产物，证明了多重PCR的特异性。向反应混合物中添加肝脏样本的细胞DNA或Q溶液对其灵敏度没有影响。通过重新检测先前经单重PCR检测呈阳性的赛鸽幼鸽肝脏样本，证明了多重PCR的实用性。
