Definition of purified enzyme-linked immunosorbent assay antigens from the culture filtrate protein of Mycobacterium bovis by proteomic analysis.
Enzyme-linked immunosorbent assay (ELISA) has been developed as the ancillary diagnosis of bovine tuberculosis at ante-mortem to overcome the disadvantages of intradermal skin test. In this study, the antigenic proteins were purified, applied to bTB ELISA, and identified through proteomic analysis. Culture filtrate protein of Mycobacterium bovis was fractionated by MonoQ column chromatography, and examined the antigenicity by immunoblotting. The antigenic 20 kDa protein was in-gel digested and identified the antigenome by LTQ mass spectrometer and peptide match fingerprinting, which were MPB64, MPB70, MPB83, Fas, Smc, Nrp, RpoC, Transposase, LeuA, and MtbE. The 20 kDa protein exhibited the highest antigenicity to bTB positive cattle in ELISA and would be useful for bTB serological diagnosis.
Cho
Yun Sang
YS
Bacteriology and Parasitology Division, National Veterinary Research Institute, National Veterinary Research & Quarantine Service, Anyang, Kyounggi, Republic of Korea. choys@nvrqs.go.kr
Lee
Sang-Eun
SE
Ko
Young Joon
YJ
Cho
Donghee
D
Lee
Hyang Shim
HS
Hwang
Inyeong
I
Nam
Hyangmi
H
Heo
Eunjung
E
Kim
Jong Man
JM
Jung
Sukchan
S
D000818:Animals; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000662:veterinary*; D009163:Mycobacterium bovis / Q000276:immunology*; D010449:Peptide Mapping / Q000379:methods / Q000662:veterinary; D012680:Sensitivity and Specificity; D014380:Tuberculosis, Bovine / Q000175:diagnosis*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins