无特定病原体鸡组织中寨卡病毒的检测及对实验室感染的血清学反应：一项比较研究

Phipps L P, Gough R E, Ceeraz Vanessa, Cox W J, Brown I H

Virology Department, VLA Weybridge, Addlestone, Surrey, UK.

Avian Pathol. 2007 Aug;36(4):301-5. doi: 10.1080/03079450701460492.

Using an isolate of West Nile virus (WNV) from lineage 1 (Goose/Israel 1998), groups of specific pathogen free chickens were experimentally infected via the subcutaneous or intravenous routes. To evaluate the relative efficiency of detecting the virus in the infected chickens, samples from a range of tissues and organs were examined by virus isolation tests in tissue culture, including Vero, primary chicken embryo liver and fibroblast cells, and polymerase chain reaction (PCR) analyses. Additionally, in order to investigate the serological response of the chickens and produce WNV monospecific antibodies, serum samples were collected from the birds during the trial and analysed for antibodies by virus neutralization (VN) and the plaque-reduction neutralization test (PRNT). No clinical signs or gross pathological changes were seen in any of the inoculated chickens throughout the study. The nested PCR used in the study appeared to be significantly more sensitive at detecting the presence of the virus in both the tissues and the inoculated Vero cell cultures compared with the detection of gross cytopathic changes as observed in infected Vero cell culture. No cytopathic changes were seen in the inoculated avian cell cultures. Following primary inoculation of the chickens there was a weak antibody response 15 days post-inoculation. However, following re-inoculation with inactivated WNV and adjuvant there was a substantial increase in the neutralizing antibody titres when tested 2 weeks later. The results obtained suggested that the PRNT was more sensitive than the conventional VN test. Based on detection of virus and serology there was no evidence of viral transmission to the close contact controls. It can be concluded that the PCR used in this study was more sensitive than virus isolation for the detection of WNV while the PRNT also appeared more sensitive than the conventional VN test.

使用来自1型西尼罗河病毒（WNV）株（1998年以色列鹅株），将特定病原体清除鸡群通过皮下或静脉途径进行实验性感染。为了评估在受感染鸡中检测病毒的相对效率，通过在包括Vero细胞、原代鸡胚肝和成纤维细胞的组织培养中进行病毒分离试验以及聚合酶链反应（PCR）分析，对一系列组织和器官的样本进行了检测。此外，为了研究鸡的血清学反应并制备WNV单特异性抗体，在试验期间从鸡中采集血清样本，并通过病毒中和（VN）和蚀斑减少中和试验（PRNT）分析抗体。在整个研究过程中，任何接种的鸡均未出现临床症状或明显的病理变化。与在受感染的Vero细胞培养物中观察到的明显细胞病变变化检测相比，该研究中使用的巢式PCR在检测组织和接种的Vero细胞培养物中病毒的存在方面似乎明显更敏感。在接种的禽类细胞培养物中未观察到细胞病变变化。鸡初次接种后，接种后15天出现微弱的抗体反应。然而，在用灭活的WNV和佐剂再次接种后，2周后检测时中和抗体滴度大幅增加。获得的结果表明，PRNT比传统的VN试验更敏感。基于病毒检测和血清学，没有证据表明病毒传播给密切接触的对照鸡。可以得出结论，本研究中使用的PCR在检测WNV方面比病毒分离更敏感，而PRNT似乎也比传统的VN试验更敏感。