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Serological Evaluation of Thin-Layer Immunoassay–Enzyme-Linked Immunosorbent Assay for Antibody Detection in Human Trichinellosis

机译:旋毛虫病抗体检测的薄层免疫分析-酶联免疫吸附测定的血清学评估

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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 μl each of antigen (7 μg/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.
机译:使用由旋毛虫(Trichinella spiralis)肌肉幼虫制备的排泄和分泌产物,评估了一种名为薄层免疫测定-酶联免疫吸附测定(TIA-ELISA)的新的免疫酶试验,以检测人旋毛虫病中的抗体。肌肉活检阳性或症状与疾病相适应的患者的血清样品(分别为 n = 8或26),在酶联免疫电转移印迹法(EITB)中均具有反应性,以及67个血清样品在ELISA和TIA-ELISA中对来自健康EITB阴性人群的样本进行了测试。通过在每个聚苯乙烯塑料培养皿中加入10μl抗原(7μg/ ml)的斑点,然后加入稀释的血清和结合物,进行TIA-ELISA。最后,加入与琼脂混合的底物以进行反应。酶副产物易于用肉眼检测为定义的点。 ELISA的灵敏度和特异性分别为76%和94%,TIA-ELISA的两个参数均为91%。两项测试与EITB的kappa相关指数分别为0.73和0.80。 TIA-ELISA可以在3小时内用普通实验室设备进行,并且使用的抗原量要比EITB和ELISA少。由于TIA-ELISA易于执行,便宜,灵敏且特异,因此该测试可用于缺乏缺乏ELISA和EITB所需专用设备的临床实验室以及人旋毛虫病抗体检测的现场研究中。

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