Staphylococcus aureus proteins differentially recognized by the ovine immune response in mastitis or nasal carriage

Serological proteome analysis of Corynebacterium pseudotuberculosis isolated from different hosts reveals novel candidates for prophylactics to control caseous lymphadenitis

“…Bacterial strains, growth conditions, and preparation of protein samples C. pseudotuberculosis 1002 and C231 strains -which have been isolated from goat and sheep, respectively -were cultivated in RPMI medium (Sigma, Saint Quentin Fallavier, France) for 24 h at 37 8C, under agitation (100 rpm). Exoproteins were extracted from the supernatant of culture in exponential growth phase (OD 0.4) using trichloroacetic acid and ethanol, as described previously (Seyffert et al, 2012). Proteins were quantified using the Bradford assay (Bradford, 1976).…”

“…Finally, the membranes were washed in ultrapure water, scanned with an Image Scanner II (Amersham Biosciences), and analyzed using Image-Quant (GE-Healthcare) software. Immune-reactive proteins were identified using Nano-ESI-MS/MS and the MASCOT software package as described previously (Seyffert et al, 2012;Le Maré chal et al, 2011). Briefly, for each protein identified in NanoLC-ESI-MS/MS, a minimum of four peptides with MASCOT score corresponding to a P value below 0.05 or an exponentially modified protein abundance index greater (Ishihama et al, 2005) than 0.4 were necessary for validation with a high degree of confidence.…”

“…However, the search for potential prophylactic candidates should also consider proteins of the pathogen proteome that are capable of stimulating the host immune response (Bhavsar et al, 2007). To this end, some studies have employed the serological proteome analysis (SERPA) to detect and identify immune-reactive proteins (i.e., proteins that are recognized by the serum of infected animals and thus produced during the infection) in search for information on pathogen biology and the host immune response to infection by bacteria (Le Maré chal et al, 2011;Seyffert et al, 2012). The SERPA approach has allowed the identification of six C. pseudotuberculosis 1002 exoproteins related to dormancy, colonization, and microbial metabolite transport .…”

Molecular characterization for some virulence and antibiotic resistance genes of Staphylococcus aureus isolated from dairy cattle's subclinical mastitis in EL- Sharkia Governorate

“…taphylococcus aureus is an opportunistic pathogen in dairy ruminant where it is found in healthy carriage and can be a major cause of mastitis (Seyffert et al, 2012). S.aureus is recognized worldwide as a frequent cause of subclinical intramammary infections in dairy cows.…”

An Optimized Fluorescence-Based Bidimensional Immunoproteomic Approach for Accurate Screening of Autoantibodies

“…In contrast to SEREX, SERPA offers the advantage of considering the post-translational modifications in the natural context of protein expression. SERPA has been extensively used in different conditions and animal models such as allergic [ 2 ] and autoimmune [ 3 – 6 ] diseases but also in other conditions: cancerology [ 7 – 9 ], metabolism [ 10 ], toxicology [ 11 ] and infectiology [ 12 – 18 ]. However, after a decade of using SERPA, some methodological problems have emerged and must be emphasized.…”