A comparison of selected immunological techniques used to detect anti-leishmanial antibodies in the sera of two reptile species

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 > 0.82) and with ELISA values for different antigen preparations compared with each other both control (r > 0.67) and immune (r > 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 of reservoirs of infective flagellates in epidemiological studies aimed at disease control, especially in areas where human infections are prevalent.