Gómez-Priego A, Crecencio-Rosales L, de-La-Rosa J L
Departamento de Zoonosis, Instituto Nacional de Diagnóstico y Referencia Epidemiológicos, Secretaría de Salud, Mexico City, Mexico.
Clin Diagn Lab Immunol. 2000 Sep;7(5):810-2. doi: 10.1128/CDLI.7.5.810-812.2000.
A new immunoenzymatic test, named the thin-layer immunoassay-enzyme-linked immunosorbent assay (TIA-ELISA), was evaluated for antibody detection in human trichinellosis using excretion and secretion products prepared from Trichinella spiralis muscle larvae. Serum samples from people with positive muscle biopsies or symptoms compatible with the disease (n = 8 or 26, respectively), all reactive in enzyme-linked immunoelectrotransfer blot assay (EITB), as well as 67 serum samples from healthy, EITB-negative people, were tested in an ELISA and TIA-ELISA. TIA-ELISA was performed in polystyrene plastic petri dishes by adding dots of 10 microl each of antigen (7 microg/ml) followed by adding diluted serum and the conjugate. Finally, the substrate mixed with agar was added to develop the reaction. Enzymatic by-products were easily detected by the naked eye as defined dots. Sensitivity and specificity were 76 and 94% for ELISA, and both parameters were 91% for TIA-ELISA. The kappa correlation indices for both tests in relation to EITB were 0.73 and 0.80, respectively. The TIA-ELISA can be carried out with common laboratory equipment in 3 h and uses lower quantities of antigen than EITB and ELISA. Since TIA-ELISA is easy to perform, cheap, sensitive, and specific, the test could be an acceptable alternative to use in clinical laboratories lacking specialized equipment needed for ELISA and EITB and in field studies for antibody detection in human trichinellosis.
一种名为薄层免疫分析 - 酶联免疫吸附测定(TIA - ELISA)的新型免疫酶检测方法,使用旋毛虫肌肉幼虫制备的排泄和分泌产物,对人体旋毛虫病中的抗体检测进行了评估。对肌肉活检呈阳性或有与该病相符症状的人群(分别为n = 8或26)的血清样本进行检测,这些样本在酶联免疫电转移印迹分析(EITB)中均呈阳性反应,同时对67份来自健康、EITB阴性人群的血清样本进行ELISA和TIA - ELISA检测。TIA - ELISA在聚苯乙烯塑料培养皿中进行,先加入10微升抗原(7微克/毫升)的点样,随后加入稀释的血清和结合物。最后,加入与琼脂混合的底物以显色反应。酶促副产物肉眼可轻易检测为明确的斑点。ELISA的灵敏度和特异性分别为76%和94%,TIA - ELISA的这两个参数均为91%。两种检测方法与EITB相关的kappa相关指数分别为0.73和0.80。TIA - ELISA可使用普通实验室设备在3小时内完成,并且比EITB和ELISA使用的抗原量更少。由于TIA - ELISA操作简便、成本低廉、灵敏且特异,该检测方法对于缺乏ELISA和EITB所需专业设备的临床实验室以及人体旋毛虫病抗体检测的现场研究而言,可能是一种可接受的替代方法。