Use of a 45 kDa protein in the detection of Campylobacter jejuni

Two techniques, both using the 45 kilodalton (kDa) outer membrane protein of Campylobacter jejuni to detect the presence of C. jejuni are described. The first technique describes a novel approach in the production of monospecific antiserum against a 45 kDa C. jejuni protein. C. jejuni was lysed, electrophoresed on polyacrylamide gel and blotted on to a polyvinyldifluoride (Immobilon-P) membrane. The membrane was stained with 0.01% Coomassie Blue, then destained to visualize protein bands. The 45 kDa band was excised from the membrane and inserted subcutaneously into chickens. After four insertions, the chickens produced a specific antibody to this protein, as determined by immunoblot. The second technique uses the same excised Immobilon-P membrane for amino acid sequence analysis. After analysis, an oligonucleotide of the corresponding sequence of the amino acids was synthesized. After labelling with an enhanced chemiluminescence kit, the oligonucleotide probe was capable of detecting the presence of C. jejuni in culture using dot-blot hybridization assays.