Tsukamoto K, Hihara H, Kono Y
Poultry Disease Laboratory, National Institute of Animal Health, Gifu, Japan.
J Vet Med Sci. 1991 Jun;53(3):399-408. doi: 10.1292/jvms.53.399.
An enzyme-linked immunosorbent assay (ELISA) for detecting avian leukosis virus (ALV) antigens was developed with rabbit anti-ALV serum. The ELISA detected purified ALV of subgroups A and B at a concentration of 0.4 ng/well and about 10(3) infectious units/well estimated by a resistance-inducing factor (RIF) test, and antigens in culture fluids from chicken embryo fibroblasts infected with subgroups A, B or E of ALV. These results showed that common antigens among the subgroups were detected by the ELISA. When virus titration was performed, virus infectivity could be determined by the ELISA within 7 days after cultivation. The titer was similar to that obtained by the RIF test on 19 days after 3 subcultures. These results indicate that the ALV-isolation test by the ELISA was superior to the RIF test in rapidity and applicability to large-scale field trials. Four specific pathogen-free (SPF) chicken lines maintained in this laboratory were examined for endogenous ALV antigens by the ELISA. Sera from laying hens had considerably high absorbance (A) values, whereas albumen samples showed low A values except for some samples (7/40 hens). Although most of sera from 1-day-old SPF chicks showed lower A values than those from laying hens, some sera showed A values as high as those from viremic chicks in 2 lines. Endogenous ALV was isolated from sera from laying hens (6/40) and their albumens (4/7) with high A values. Two SPF chicken lines were found to produce endogenous virus at a high frequency.
用兔抗禽白血病病毒（ALV）血清建立了一种检测ALV抗原的酶联免疫吸附试验（ELISA）。该ELISA能检测到浓度为0.4 ng/孔的A和B亚群纯化ALV，以及通过抗性诱导因子（RIF）试验估计的约10³感染单位/孔，还能检测感染ALV A、B或E亚群的鸡胚成纤维细胞培养液中的抗原。这些结果表明该ELISA能检测到亚群间的共同抗原。进行病毒滴定时，培养后7天内可通过ELISA确定病毒感染性。其滴度与传代3次后19天通过RIF试验获得的滴度相似。这些结果表明，ELISA法进行ALV分离试验在速度和适用于大规模现场试验方面优于RIF试验。用该ELISA检测了本实验室饲养的4个特定病原体-free（SPF）鸡品系的内源性ALV抗原。产蛋母鸡血清的吸光度（A）值相当高，而蛋清样本除部分样本（7/40只母鸡）外A值较低。虽然1日龄SPF雏鸡的大多数血清A值低于产蛋母鸡，但2个品系中一些血清的A值与病毒血症雏鸡的一样高。从A值高的产蛋母鸡血清（6/40）及其蛋清（4/7）中分离出了内源性ALV。发现2个SPF鸡品系高频产生内源性病毒。
