Evaluation of Recombinant Proteins Based Indirect ELISA and Microscopic Agglutination Test for Seroprevalence Study of Feline Leptospirosis
نویسندگان
چکیده
Leptospirosis is worldwide zoonotic disease that is serious problem in south Indian states and now emerging in North India. The focus these days is to develop recombinant outer membrane protein based Enzyme-linked immunosorbent assay (ELISA) for seroprevalence of leptospirosis. The aim of this study was to evaluate indirect ELISA based on recombinant LipL41, Loa22, LipL21, LipL32 and OmpL1 proteins in comparison with Microscopic agglutination test (MAT). For this His-tagged recombinant proteins were expressed in DH5α host cells and purified by Ni-NTA affinity chromatography. Hyper-immune sera raised against recombinant proteins in rabbits confirmed by dot blotting and appearance of dark dot confirm the successful antigen-antibody binding. Leptospirosis suspected 7 tiger and 3 lion sera samples screened with rLipL41+Loa22+LipL21and rLipL41+LipL32+OmpL1antigen combinations based indirect ELISA. Only 2 tiger samples were positive by MAT against Grippotyphosa and Icterohemorrhagae serovars in comparison to 6 sera samples positive by rLipL41 +Loa22 + LipL21 and rLipL41 +LipL32 + OmpL1antigen combination based indirect ELISA. Out of 3 lion samples 2 were positive by MAT against Icterohemorrhagae serovar while in rLipL41 +Loa22 + LipL21 and rLipL41 +LipL32 + OmpL1 antigen combination based indirect ELISA all were positive. MAT titre values were lower than indirect ELISA titre values revealed that ELISA is more sensitive than MAT. rLipL41+LipL32+OmpL1 antigen combinations based indirect ELISA showed high titre values as compared to rLipL41+Loa22+LipL21antigen combination based ELISA that confirmed the presence of most immunodominant protein LipL32 in the combination.
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