Immunodiagnosis of Besnoitia besnoiti infection by ELISA and Western blot

A longitudinal study of Besnoitia besnoiti infections and seasonal abundance of Stomoxys calcitrans in a dairy cattle farm of southwest France

“…A combination of serological analyses (Western blots and ELISA), clinical examinations and entomological survey was used. The Western blot, adapted from Cortes et al (2006a), had been published to have a sensitivity of 91.3% and a specificity of 96.4%, based upon assessment with parasitologically and clinically manifest/proven cases. According to the manufacturer, the sensitivity and the specificity of the PrioCHECK ® Besnoitia Ab ELISA are both around 98%, also based primarily upon testing of clinical cases.…”

“…Nine animals (15.8%) showed SCC, 31 (54.4%) were seropositive without any sign of infection and 15 (26.3%) were negative in serology. According to Cortes et al (2006a) and Schares et al (2010), some animals with clinical signs of besnoitiosis remain serologically negative. We did not observe this subset in our survey.…”

“…Antigen extraction and Western blot (adapted from Cortes et al, 2006a) Infected Vero cells and free tachyzoites were harvested and passed through a 25-gauge needle to release parasites. They were separated on a Sephadex G-25 column (GE Healthcare Europe Gmbh, Ref 17-0851-01, Orsay, France) and centrifuged at 1200 × g for 10 min at 4 • C. Pellets of tachyzoites were frozen at −80 • C until use.…”

“…Electrophoresis was performed in SDS-PAGE (12.5%, w/v) gels using 15 l sample/cm of slot. Then, B. besnoiti antigens were electrophoretically transferred to PVDF membranes Cortes et al, 2006a): domain I ranging between 12 and 20 kDa, domain II ranging between 23 and 38 kDa and domain III ranging between 60 and 90 kDa with at least a set of four bands within each of them. This Western blot (WB) technique exhibited a diagnostic sensitivity of 91.3% and a specificity ranging between 96.4% and 100% (Cortes et al, 2006a).…”

Besnoitia besnoiti protein disulfide isomerase (BbPDI): Molecular characterization, expression and in silico modelling

“…In Europe, besnoitiosis has been recently reported in Portugal (Cortes et al, 2005(Cortes et al, , 2006a, Spain (Castillo et al, 2009;Fernandez-Garcia et al, 2010;Irigoien et al, 2000), France (Alzieu et al, 2007;Bourdeau et al, 2004;Lienard et al, 2010), Italy (Agosti et al, 1994;Gollnick et al, 2010) and Germany (Mehlhorn et al, 2009;Schares et al, 2009). There is no treatment against this infection despite the efforts to identify and develop effective agents (Cortes et al, 2007;Shkap et al, 1987).…”