Buscaglia C, Bankowski R A, Miers L
Avian Dis. 1985 Jul-Sep;29(3):672-80.
Two serological tests--the virus-neutralization (VN) test in chicken embryo fibroblasts (CEF) using a cell-culture-adapted virus, and the enzyme-linked immunosorbent assay (ELISA)--were used for evaluating the immune response in chickens against fowlpox virus. The VN test was conducted in 96-well tissue-culture plates using a fowlpox virus that was adapted to induce cytopathic effects (CPE) in CEF in 48 hr. The ELISA was carried out with an antigen prepared by precipitation of a cell-culture-propagated virus suspension with ammonium sulfate and concentration by centrifugation. A 0.1 M acetate buffer, pH 5, was used as the sensitizing solution for maximum specific binding of the antigen to the microplate plastic well. No antibodies were detected by the VN test in 228 serum samples taken from chickens at irregular intervals between 1 and 39 weeks of age, even though the birds were vaccinated against fowlpox at 13 weeks of age. However, in sera collected 4 weeks after a sample of laying hens was challenged with fowlpox virus, VN titers of 1/10 to 1/40 were detectable. On the other hand, significant antibody reactions were detected by the ELISA on sera from chickens during the growing period, following vaccination and challenge. Although no maternal antibodies were found at 1 week of age, a continuous increase in the mean ELISA titers to fowlpox was demonstrated during the entire experimental period. This study showed that the ELISA was considerably more sensitive and practical than the VN test.
两种血清学检测方法——使用适应细胞培养的病毒在鸡胚成纤维细胞（CEF）中进行的病毒中和（VN）试验，以及酶联免疫吸附测定（ELISA）——被用于评估鸡对禽痘病毒的免疫反应。VN试验在96孔组织培养板中进行，使用一种能在48小时内在CEF中诱导细胞病变效应（CPE）的禽痘病毒。ELISA则使用通过用硫酸铵沉淀细胞培养繁殖的病毒悬液并通过离心浓缩制备的抗原进行。使用pH值为5的0.1M醋酸盐缓冲液作为致敏溶液，以使抗原与微孔板塑料孔最大程度地特异性结合。在1至39周龄期间不定期采集的228份鸡血清样本中，通过VN试验未检测到抗体，尽管这些鸡在13周龄时接种了禽痘疫苗。然而，在一组产蛋母鸡用禽痘病毒攻击后4周采集的血清中，可检测到1/10至1/40的VN滴度。另一方面，在生长期间、接种疫苗后和攻击后，ELISA在鸡的血清中检测到了显著的抗体反应。虽然在1周龄时未发现母源抗体，但在整个实验期间，ELISA对禽痘的平均滴度持续升高。这项研究表明，ELISA比VN试验灵敏得多且更实用。
