Tadese Theodros, Potter Andy E, Fitzgerald Scott, Reed Willie M
Department of Pathobiology and Diagnostic Investigation and Diagnostic Center for Population and Animal Health, Michigan State University, East Lansing, MI 48824, USA.
Avian Dis. 2007 Sep;51(3):719-24. doi: 10.1637/0005-2086(2007)51[719:CIICWF]2.0.CO;2.
A concurrent infection of chickens with infectious laryngotracheitis virus (ILTV), a herpesvirus, and fowlpox virus (FWPV), an avipoxvirus, is described. Two techniques, an immunohistochemistry (IHC) technique and a multiplex polymerase chain reaction (PCR), were used to examine 11 tissue samples from chickens clinically diagnosed as FWPV-infected, but only IHC was used to examine six tissue-paraffin blocks prepared from turkeys suspected of having FWPV infection. By multiplex PCR, both FWPV and ILTV were detected from three chicken samples (FI-90, FI-93, and FI-94); both FWPV and ILTV were detected from only two samples (FI-93 and FI-94) by IHC. All chicken samples were positive for FWPV by both PCR and IHC. Viral DNA from these samples was further confirmed by restriction enzyme analysis. When turkey samples were analyzed by the double-stain IHC, all six samples showed the presence of FWPV antigens, but no ILTV antigens. The double IHC technique, using monoclonal antibodies against FWPV and ILTV, was successful in simultaneous demonstration of specific FWPV and ILTV antigens colocalized in infected tissue samples as well as within individual cells. This paper emphasizes the importance of reliable tests that detect specifically the presence of ILTV and FWPV in infected tissue samples. The multiplex PCR assay holds potential to be versatile, rapid, and more sensitive (100%) than IHC (67%) for the simultaneous detection of two different avian viruses. Furthermore, the presence of mixed infection should always be kept in mind in the virologic analysis of respiratory sickness of poultry.
本文描述了鸡同时感染传染性喉气管炎病毒（ILTV，一种疱疹病毒）和禽痘病毒（FWPV，一种禽痘病毒）的情况。采用两种技术，即免疫组织化学（IHC）技术和多重聚合酶链反应（PCR），对11份临床诊断为FWPV感染的鸡组织样本进行检测，而对于6份怀疑感染FWPV的火鸡组织石蜡块仅采用IHC技术检测。通过多重PCR，在3份鸡样本（FI - 90、FI - 93和FI - 94）中检测到FWPV和ILTV；通过IHC仅在2份样本（FI - 93和FI - 94）中检测到FWPV和ILTV。所有鸡样本通过PCR和IHC检测FWPV均呈阳性。这些样本的病毒DNA通过限制性酶切分析进一步得到证实。当对火鸡样本进行双重染色IHC分析时，所有6份样本均显示存在FWPV抗原，但未检测到ILTV抗原。使用针对FWPV和ILTV的单克隆抗体的双重IHC技术成功地同时显示了在感染组织样本以及单个细胞中共定位的特异性FWPV和ILTV抗原。本文强调了可靠检测在特异性检测感染组织样本中ILTV和FWPV存在的重要性。多重PCR检测对于同时检测两种不同禽病毒具有通用性、快速性且比IHC（67%）更灵敏（100%）的潜力。此外，在禽类呼吸道疾病的病毒学分析中应始终牢记混合感染的存在。
