Hopkins B A, Skeeles J K, Houghten G E, Story J D
Department of Animal Sciences, University of Arkansas, Fayetteville 72701.
Avian Dis. 1988 Apr-Jun;32(2):353-61.
A Bordetella avium enzyme-linked immunosorbent assay (ELISA) was developed to detect serum antibodies in 1-day-old poults, experimentally infected turkeys, and naturally infected turkeys. The optimized procedure included use of a suspension of whole bacteria coated onto plastic microtiter plates, a 1:200 serum dilution, a 1:3200 dilution of commercially available goat anti-turkey IgG (heavy and light chain) conjugated with horseradish peroxidase, and 0.04% orthophenylenediamine as substrate. A sample/negative (S/N) ratio method of analysis was used to estimate antibody titer from absorbance values. The regression equation used to estimate antibody titers was derived from the testing of naturally infected turkey sera. The equation was derived by plotting the log10 titer of the sera against the S/N ratio at a 1:200 serum dilution. The ELISA was an effective method for detecting antibody to B. avium, and the procedure should prove useful for laboratories equipped for high-volume ELISA testing.
开发了一种禽博德特氏菌酶联免疫吸附测定（ELISA），用于检测1日龄雏火鸡、实验感染火鸡和自然感染火鸡血清中的抗体。优化后的步骤包括使用包被在塑料微量滴定板上的全菌悬液、1:200的血清稀释度、1:3200稀释的与辣根过氧化物酶结合的市售山羊抗火鸡IgG（重链和轻链），以及0.04%的邻苯二胺作为底物。采用样本/阴性（S/N）比值分析方法，根据吸光度值估算抗体滴度。用于估算抗体滴度的回归方程源自对自然感染火鸡血清的检测。该方程是通过将血清的log10滴度与1:200血清稀释度下的S/N比值作图得出的。ELISA是检测禽博德特氏菌抗体的有效方法，该方法对具备大量ELISA检测能力的实验室应具有实用价值。
