Watthanakulpanich Dorn, Waikagul Jitra, Dekumyoy Paron, Anantaphruti Malinee T
Department of Helminthology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Southeast Asian J Trop Med Public Health. 2003;34 Suppl 2:114-20.
Antigens derived from somatic extracts of Bithynia funiculata, an intermediate snail host of O. viverrini, have been demonstrated to be highly heterogeneous in molecular weight (MW). These antigens have been suggested to be of potential use for serodiagnosis. In this study, B. funiculata somatic antigens were extracted using five different centrifugal speeds, namely 10,000 (C1); 20,000 (C2); 30,000 (C3); 40,000 (C4) and 50,000 (C5) rpm, with the aim of removing some non-specific antigens and determining the optimal centrifugal speed to obtain the highest efficiency of the test for which they will be used. The enzyme-linked immunosorbent assay technique was used to compare the reactivity of the five different centrifugal speed-prepared antigens. The sensitivity and specificity of all five antigens were compared by testing against sera from 81 opisthorchiasis patients, 30 parasite-free healthy individuals and 50 individuals infected with other helminthic infections, using mean + 4SD of all healthy individuals as the cut-off value. The sensitivity of these antigens was 69.1, 84.0, 80.2, 84.0 and 70.4, respectively; while the specificity was 66.2, 76.2, 82.5, 86.2 and 71.2, respectively. Immunoreactive components of each antigen were analyzed by SDS-PAGE and Western blot technique. The assay showed that three pairs of antigens with MW of 29 and 30, 47 and 50, and 86 and 90 kDa of all five antigens, which have previously been shown to be highly immunogenic, still reacted with a pooled serum from patients with opisthorchiasis. However, the C4 antigens gave more distinct components. Our results showed that 40,000 rpm is the optimal speed for antigen preparation for use in the serological diagnosis of opisthorchiasis, as demonstrated by the most satisfactory results of both sensitivity and specificity in the indirect ELISA and Western blot technique.
来自猫后睾吸虫中间螺宿主缢蛏（Bithynia funiculata）虫体提取物的抗原，已被证明在分子量（MW）上具有高度异质性。这些抗原已被认为具有用于血清学诊断的潜在用途。在本研究中，使用五种不同的离心速度，即10,000（C1）；20,000（C2）；30,000（C3）；40,000（C4）和50,000（C5）转/分钟，提取缢蛏虫体抗原，目的是去除一些非特异性抗原，并确定能获得最高检测效率的最佳离心速度，以便用于后续检测。采用酶联免疫吸附测定技术比较了五种不同离心速度制备的抗原的反应性。以所有健康个体的平均值 + 4标准差作为临界值，通过检测81例华支睾吸虫病患者、30例无寄生虫感染的健康个体和50例感染其他蠕虫的个体的血清，比较了所有五种抗原的敏感性和特异性。这些抗原的敏感性分别为69.1、84.0、80.2、84.0和70.4；而特异性分别为66.2、76.2、82.5、86.2和71.2。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）和蛋白质免疫印迹技术分析了每种抗原的免疫反应成分。检测显示，所有五种抗原中分子量为2
