Development of a double-antibody elisa for IgG evaluation in pichis (Zaedyus pichiy, Xenarthra, Dasypodidae) with special interest for epidemiological surveillance.

MORENO-SOSA MT; ACTIS EA; PENNACCHIO GE; JAHN GA; MACKERN OBERTI JP; SUPERINA ME

Mendoza

Congreso; XXXIV Reunión Científica Anual de la Sociedad de Biología de Cuyo; 2016

Sociedad de Biología de Cuyo

Many zoonotic pathogens have a wildlife reservoir, and hunters are especially prone to acquire a zoonotic disease while killing or consuming wildlife. The pichi (Zaedyus pichiy ; Xenarthra, Dasypodidae) is an intensely hunted armadillo species. Although several biological features of the pichi are well studied, data about its potential role as a target or a reservoir of human pathogens, such as Trypanosoma cruzi and Brucella abortus , are scarce. Due to the lack of commercially available serological tests to detect specific IgG, the aim of this work was to develop a serological test to evaluate the presence in pichis of IgG specific to zoonotic pathogens important to public health. To this end we developed a double-antibody indirect ELISA. Gamma heavy chains (≈55kDa) from pichi serum were purified through SDS-PAGE and used to immunize BALB/c mice with Freund complete adjuvant and mice serum samples were collected. To confirm the specificity to pichi IgG of the antiserum we performed WB with pichi serum proteins. WB membranes were first incubated with antiserum from immunized mice, and secondarily incubated with goat anti mouse IgG-HRP. After detection, lanes containing pichi serum displayed only a band between 50-60kDa which correspond to pichi gamma heavy chains. Similarly, we found that our mouse antiserum showed reactivity against purified pichi gamma heavy chains by ELISA. In conclusion, these data suggest that we have developed a useful tool to evaluate specific IgG levels from pichis. This antiserum is crucial to indirectly evaluate whether pichis have been infected or been in contact with several pathogens, including the zoonotic parasite Trypanosoma cruzi.