Wirtz R A, Charoenvit Y, Burkot T R, Esser K M, Beaudoin R L, Collins W E, Andre R G
Department of Entomology, Walter Reed Army Institute of Research, Washington, D.C. 20307-5100.
Med Vet Entomol. 1991 Jan;5(1):17-22. doi: 10.1111/j.1365-2915.1991.tb00515.x.
Nine monoclonal antibodies (MAbs) developed against Plasmodium vivax (Grassi & Feletti) salivary gland sporozoites were evaluated for use in an enzyme-linked immunosorbent assay (ELISA), using sporozoites developed in Anopheles dirus Peyton & Harrison An. gambiae Giles and An.maculatus Theobald. Four of the antibodies were unsuitable due to the low sensitivity of the resulting assays or the requirement for high concentrations of capture antibody. An additional two MAbs were rejected because they resulted in assays with high background absorbance, attributed to self-binding. Of the three remaining MAbs, the use of Navy vivax sporozoite (NVS) 3 resulted in an ELISA with the highest sensitivity and the lowest concentration requirement for capture antibody. Assay sensitivity varied with sporozoite strain indicating possible quantitative epitope heterogeneity. None of the MAbs cross-reacted with the heterologous sporozoites tested by immunofluorescence antibody assay (IFA). The IFA activity was not an indicator of ELISA sensitivity. The use of MAb NVS 3 in a standardized ELISA method resulted in an assay 10 times more sensitive than reported previously for P. vivax sporozoites, with a detection limit of fewer than 100 sporozoites per mosquito.
针对间日疟原虫（Grassi & Feletti）唾液腺子孢子研制的9种单克隆抗体（MAb），采用在大劣按蚊（Peyton & Harrison）、冈比亚按蚊（Giles）和多斑按蚊（Theobald）体内发育的子孢子，对其在酶联免疫吸附测定（ELISA）中的应用进行了评估。由于所得测定的灵敏度低或对捕获抗体的浓度要求高，4种抗体不适用。另外2种单克隆抗体也被排除，因为它们导致测定的背景吸光度高，这归因于自身结合。在剩下的3种单克隆抗体中，使用间日疟原虫子孢子（NVS）3可得到灵敏度最高且捕获抗体浓度要求最低的ELISA。测定灵敏度随子孢子株而变化，表明可能存在定量表位异质性。通过免疫荧光抗体测定（IFA）测试，没有一种单克隆抗体与异源子孢子发生交叉反应。IFA活性不是ELISA灵敏度的指标。在标准化ELISA方法中使用单克隆抗体NVS 3，得到的测定比先前报道的间日疟原虫子孢子测定灵敏度高10倍，检测限为每只蚊子少于100个子孢子。
