The specificity of the developed antibodies was studied by the assessment of binding affinity of heat-killed bacterial strains to antibodies immobilized in the SPR system (Table 1). The SPR analysis of ulcer secretions using biosensor chips immobilized with antibodies developed against P. aeruginosa, S. aureus, E. fecalis, and S. epidermidis determined that S. aureus was found in 6/10 cultures from chronic ulcers and there was binding affinity to antibodies against S. aureus in 5/6 SPR analyses. S. aureus grew in 2/5 ulcer secretions from patients that had undergone breast cancer surgery and there was a binding affinity to S. aureus antibodies in both cases. E. fecalis grew in 1/10 ulcer secretion from chronic ulcers, however there was a positive binding to antibodies against E. fecalis in 3/10 specimens. There was no binding affinity to antibodies against E. fecalis in the ulcer secretions from healthy staff or patients who underwent breast cancer surgery (Table 2).Table 1Specificity test of in vitro antibodiesControlsAnti-S. aureus Anti-S. epidermidis Anti-E. fecalisAnti-P. aeruginosa Anti-C. perfringens Anti- B. fragilis Anti-P. oralis Anti-E. coli Anti-Streptococcus sp. S. aureus 25000000000 S. epidermidis 01030000000 E. fecalis 00244000000 P. aeruginosa 00036.400000 C.perfringens 000031.80000 B. fragilis 00000230.1000 P. oralis 00000057000 E. coli 0000000185.50 Streptococcus sp. 00000000278.3Anti-guinea pig IgG000000000Anti-human IgG279140646.5439342.375.8809.2234.6461.1Table 2Affinity of ulcer secretions to the immobilised antibodies in SPR system and the correlation to culture resultsUlcer secretionCulture resultsBinding to the produced antibodies in SPR systemAnti-P. aeruginosa Anti-S. aureus Anti-E. fecalis Anti-S. epidermidis Ulcer secretion from patients with chronic ulcer1 Staphylococcus aureus + Enterobacter cloacae negativepositivenegativepositive2 Staphylococcus aureus + Echerichia coli negativepositivenegativenegative3 Staphylococcus aureus + Pseudomonas aeroginosa negativepositivenegativenegative4 Staphylococcus aureus negativenegativenegativenegative5 Proteus mirabilis + coagulase negative staphylococcus negativenegativepositivepositive6negativenegativenegativenegativenegative7 Pseudomonas aeruginosa + Enterococcus fecalis positivenegativepositivenegative8 Staphylococcus aureus negativepositivenegativepositive9 Staphylococcus aureus negativepositivepositivepositive10negativenegativenegativenegativenegativeUlcer secretion from patients post breast cancer surgery1negativenegativenegativenegativepositive2negativenegativenegativenegativepositive3negativenegativenegativenegativepositive4 Staphylococcus aureus negativepositivenegativenegative5 Staphylococcus aureus negativepositivenegativenegativeUlcer secretion from healthy volunteers post skin biopsy in fore-arm1negativenegativenegativenegativepositive2negativenegativenegativenegativenegative3negativenegativenegativenegativenegative4negativenegativenegativenegativepositive5negativenegativenegativenegativepositive6negativenegativenegativenegativepositive7negativenegativenegativenegativepositive8negativenegativenegativenegativepositive9negativenegativenegativenegativepositive