Ingram G A, Molyneux D H
J Immunol Methods. 1984 Dec 14;75(1):53-64. doi: 10.1016/0022-1759(84)90224-2.
European green lizards (Lacerta viridis) and spiny-tailed agamids (Agama caudospinosum) were obtained from areas endemic for human leishmaniasis. Serum antibody titres against Leishmania agamae, a reptilian leishmanial species, in normal lizards and lizards injected with Leishmania agamae promastigotes were measured by 5 immunological methods commonly used in the serodiagnosis of the human and mammalian leishmaniasis viz. immobilisation test (IMM), direct agglutination (DA), complement-fixation test (CFT), indirect haemagglutination (IHA) and enzyme-linked immunosorbent assay (ELISA). Correlation coefficients (r) were determined for comparisons between each method and linear regression equations calculated to convert antibody titres by one method to those by another. In each lizard species, the IMM test gave the lowest values while the highest were obtained with ELISA. The highest mean titre obtained by ELISA was between 2 and 10 times that obtained by the other methods for both control and immune sera. The methods of preparing the leishmanial antigen extracts affected the IHA and ELISA titres, while the source of complement was critical in obtaining good CFT values. Correlations ranging from 3% to 77% were found for the control animals but higher values ranging from 65% to 96% were obtained with the immunised lizards. Overall, the best correlation was with IHA and ELISA (r greater than 0.82) and with ELISA values for different antigen preparations compared with each other for both control (r greater than 0.67) and immune (r greater than 0.90) sera. ELISA thus appears the most sensitive method for detection and quantitation of anti-flagellate antibodies in normal lizard serum and for the determination of titres in immune serum. ELISA is the most applicable technique for screening reptiles and other lower vertebrates for anti-parasite immunoglobulins, and for screening potential carriers or reservoirs of infective flagellates in epidemiological studies aimed at disease control, especially in areas where human infections are prevalent.
欧洲绿蜥（绿蜥蜴）和刺尾鬣蜥取自人类利什曼病的流行地区。采用常用于人类和哺乳动物利什曼病血清诊断的5种免疫学方法，即固定试验（IMM）、直接凝集试验（DA）、补体结合试验（CFT）、间接血凝试验（IHA）和酶联免疫吸附测定（ELISA），检测正常蜥蜴以及注射了利什曼原虫前鞭毛体的蜥蜴血清中针对蜥蜴利什曼原虫（一种爬行动物利什曼原虫）的抗体滴度。确定了每种方法之间比较的相关系数（r），并计算了线性回归方程，以便将一种方法的抗体滴度转换为另一种方法的抗体滴度。在每种蜥蜴物种中，IMM试验得到的值最低，而ELISA得到的值最高。对于对照血清和免疫血清，ELISA获得的最高平均滴度是其他方法获得的平均滴度的2至10倍。制备利什曼原虫抗原提取物的方法影响IHA和ELISA滴度，而补体来源对于获得良好的CFT值至关重要。对照动物的相关系数在3%至77%之间，但免疫蜥蜴的相关系数更高，在65%至96%之间。总体而言，最佳相关性存在于IHA和ELISA之间（r大于0.82），并且对于对照血清（r大于0.67）和免疫血清（r大于0.90），不同抗原制剂的ELISA值之间相互比较也存在最佳相关性。因此，ELISA似乎是检测和定量正常蜥蜴血清中抗鞭毛虫抗体以及测定免疫血清中抗体滴度的最灵敏方法。ELISA是筛查爬行动物和其他低等脊椎动物抗寄生虫免疫球蛋白，以及在旨在疾病控制的流行病学研究中筛查感染性鞭毛虫潜在携带者或储存宿主的最适用技术，特别是在人类感染流行的地区。
