Ruitenberg E J, Steerenberg P A, Brosi B J, Buys J
Tijdschr Diergeneeskd. 1976 Jan 15;101(2):57-70.
An enzyme innunoassay using horse radish peroxidase as teh marker enzyme for the detection of antibodies to Trichinella spiralis in pigs was described. In the enzyme-linked immunosorbent assay (ELISA) quantitation os specific antibodies is obtained by means of peroxidase labeled anti-species-immunoglobulin in antigen-coated tubes. The enzyme remaining in the tube after wasging provides a measure of the amount of specific antibodies in the serum. A crude saline extract of T. spiralis muscle larvae served as antigen. In experiments in which conventionally raised pigs intended for slaughter were infected with various numbers of T. spiralis larvae (25,000; 12,500; 5,000; 2,500; 1,500; 500; 150; 50) a positive correlation was found between initial dose of larvae administered and amount of antibodies detected by ELISA. Compared with immuno-fluorescence (IF) ELISA was more sensitive. IF yielded positive results in 11 out of 34 infected animals, whereas ELISA results were positive in 27 cases. In order to evaluate ELISA results under practical conditions extinction values os sera from both infected and non-infected conventional pigs were compared with the highest extinction value of a group of 74 negative conventional pig sera. Due to the relatively high background reaction of some of these negative sera the number of positive practical ELISA results decreased from 27 to 19 out of 34 animals. Furthermore, in 1 out of 10 non-infected animals a false positive practical ELISA result was obtained. Ways to improve the reliability of ELISA are discussed.
描述了一种使用辣根过氧化物酶作为标记酶的酶免疫测定法,用于检测猪体内旋毛虫抗体。在酶联免疫吸附测定(ELISA)中,通过在抗原包被的试管中使用过氧化物酶标记的抗种属免疫球蛋白来定量特异性抗体。洗涤后留在试管中的酶提供了血清中特异性抗体量的度量。旋毛虫肌肉幼虫的粗盐水提取物用作抗原。在将常规饲养用于屠宰的猪感染不同数量旋毛虫幼虫(25,000;12,500;5,000;2,500;1,500;500;150;50)的实验中,发现接种幼虫的初始剂量与ELISA检测到的抗体量之间呈正相关。与免疫荧光法(IF)相比,ELISA更灵敏。IF在34只感染动物中的11只中产生阳性结果,而ELISA结果在27例中呈阳性。为了在实际条件下评估ELISA结果,将感染和未感染的常规猪血清的吸光度值与一组74只阴性常规猪血清的最高吸光度值进行比较。由于其中一些阴性血清的背景反应相对较高,实际ELISA阳性结果的数量从34只动物中的27只降至19只。此外,在10只未感染的动物中有1只获得了ELISA实际假阳性结果。讨论了提高ELISA可靠性的方法。