Viljoen G J, Vermeulen N M, Neitz A W
Department of Biochemistry, Veterinary Research Institute, Onderstepoort.
Onderstepoort J Vet Res. 1987 Sep;54(3):305-12.
Numerous parameters affect the enzyme-linked immunosorbent assay activity and the assay conditions must therefore be carefully controlled to obtain reproducible results. These parameters include temperature, pH, ionic strength, buffer composition, cofactors, substrate depletion, product inhibition, increasing reversal of reaction as product concentration increases, adsorption of enzyme to solid supports, denaturation and non-enzymatic background rate. An ELISA was used to detect Cowdria ruminantium antibodies during the course of heartwater disease. IgM antibodies reached a maximum on the 4th day after infection and disappeared on the 7th day. IgG antibodies first appeared on the 8th day and continued to increase during the remainder of the observation period of 28 days. The presence of C. ruminantium in the blood fractions of diseased animals was demonstrated by an enzyme-linked immunosorbent assay. The earliest detection of C. ruminantium antigen was in plasma and serum on the 4th day after inoculation. Of all the blood fractions investigated, the red blood cell fraction showed the highest concentration and this reached a maximum on the 12th day after infection.
众多参数会影响酶联免疫吸附测定的活性，因此必须严格控制测定条件以获得可重复的结果。这些参数包括温度、pH值、离子强度、缓冲液成分、辅因子、底物消耗、产物抑制、随着产物浓度增加反应逆转加剧、酶吸附到固相载体上、变性以及非酶促背景速率。在牛心水病病程中，采用酶联免疫吸附测定法检测反刍动物考德里氏体抗体。IgM抗体在感染后第4天达到峰值，第7天消失。IgG抗体在第8天首次出现，并在为期28天的观察期剩余时间内持续增加。通过酶联免疫吸附测定法证实患病动物血液组分中存在反刍动物考德里氏体。最早检测到反刍动物考德里氏体抗原是在接种后第4天的血浆和血清中。在所研究的所有血液组分中，红细胞组分浓度最高，且在感染后第12天达到峰值。
