Bień J
Instytut Parazytologii im. Witolda Stefańiskiego PAN, ul. Twarda 51/55, 00-818 Warszawa.
Wiad Parazytol. 2006;52(3):205-12.
ELISA can be used to measure produced antibodies or Trichinella spp. antigens in the samples. They are detected with antibodies linked to an enzyme that reacts with a substrate and generate a colour reaction. The optical density (OD) of the reaction is measured spectrophotometrically. ELISA assays can be done in several different procedures called "direct", "indirect", "sa ndwich", and "competition" ELISA. Since the 1970s, the studies have been done on improving or replacing direct methods of Trichinella diagnosis with serological methods based on the ELISA. When somatic antigens of L1 T. spiralis were used, the specificity of the ELISA was poor due to a high probability of cross-reactions with other pathogens. During the 1980s the specificity of the ELISA was improved by excretory-secretory (E/S) antigens obtained during Trichinella muscle larvae incubation in vitro. Recently a synthetic glycan antigen has been developed and the increasing of ELISA specificity and sensitivity was noticed. The sensitivity of the ELISA using an E/S antigen ranging from 93.1 to 99.2% but the specificity from 90.6 to 99.4%. The ELISA method is relatively simple to apply, reliable, readily standardized and provides an acceptable balance of sensitivity and specificity. But all modified procedures should be validated. In Poland, the studies on the usefulness of ELISA for antibodies detection against T. spiralis in pigs and wild animals are limited. Own ELISA procedure was prepared in Pathophysiology Lab. in W. Stefański Institute of Parasitology of PAS. ELISA was used to examine IgG level against L1 T. spiralis in pigs and wild boars serum samples. Of 1474 pig samples, only 12 were positive. Of 1880 wild boar samples only 14 were positive. The results of this study are comparable with performance obtained using commercial sets. The results showed the usefulness of ELISA for T. spiralis diagnosis in pigs and wild boars and confirmed the possibility of use the ELISA test for application in the slaughterhouse.
酶联免疫吸附测定(ELISA)可用于检测样本中产生的抗体或旋毛虫属抗原。它们通过与一种酶相连的抗体进行检测,该酶与底物发生反应并产生颜色反应。通过分光光度法测量反应的光密度(OD)。ELISA检测可通过几种不同的程序进行,称为“直接”“间接”“夹心”和“竞争”ELISA。自20世纪70年代以来,人们一直在研究用基于ELISA的血清学方法改进或取代旋毛虫诊断的直接方法。当使用旋毛形线虫L1期的体抗原时,由于与其他病原体发生交叉反应的可能性很高,ELISA的特异性较差。在20世纪80年代,通过旋毛虫肌肉幼虫体外培养获得的排泄-分泌(E/S)抗原提高了ELISA的特异性。最近开发了一种合成聚糖抗原,并注意到ELISA的特异性和敏感性有所提高。使用E/S抗原的ELISA敏感性在93.1%至99.2%之间,而特异性在90.6%至99.4%之间。ELISA方法应用相对简单、可靠、易于标准化,并且在敏感性和特异性之间提供了可接受的平衡。但所有改进的程序都应经过验证。在波兰,关于ELISA在检测猪和野生动物中抗旋毛虫抗体的实用性研究有限。波兰科学院寄生虫学研究所W. 斯特凡斯基病理生理学实验室制备了自己的ELISA程序。ELISA用于检测猪和野猪血清样本中抗旋毛形线虫L1期的IgG水平。在1474份猪样本中,只有12份呈阳性。在1880份野猪样本中,只有14份呈阳性。本研究结果与使用商业试剂盒获得的性能相当。结果表明ELISA在猪和野猪旋毛虫诊断中的实用性,并证实了ELISA检测在屠宰场应用的可能性。