Characterization and application of a new monoclonal antibody with high specificity for Helicobacter hepaticus.
OBJECTIVE
Infection with Helicobacter hepaticus is suggested to play a role in the pathogenesis of chronic liver disease in humans. However, reactive antigens among Helicobacter species make the development of an H. hepaticus ELISA test with high specificity difficult. A new monoclonal antibody from a hybridoma clone (HRII-51) showed high specificity to H. hepaticus without cross-reaction to other gastrointestinal bacteria.


METHODS
The molecular weight of HRII-51 immunoreactive antigen was examined by Western blot of H. hepaticus probed with the monoclonal antibody HRII-51. A HRII-51-immunoreactive antigen capture ELISA was prepared in which the specific antigen was anchored by HRII-51-immobilized ELISA plate. Accuracy of HRII-51 antigen capture ELISA was examined using sera obtained from mice inoculated with Helicobacter species. Specificity of HRII-51 antigen capture ELISA was compared to that of H. hepaticus antigen-based ELISA using human sera with absorption by H. pylori cell lysate.


RESULTS
HRII-51 immunoreactive antigen had a molecular weight of 15 kDa. Sensitivity and specificity of HRII-51 antigen capture ELISA were 87.0% and 97.6% in mice inoculated with Helicobacter species. In human sera, modification of the results by absorption with H. pylori lysate was smaller in HRII-51 antigen capture ELISA comparing with H. hepaticus-antigen-based ELISA.


CONCLUSIONS
Use of the HRII-51 antigen capture ELISA would be a useful approach for the serodiagnosis of H. hepaticus infection in both experimental animals and humans.
Fukuda
Yoshihiro
Y
Hyōgo College of Medicine, Nishinomiya, Japan.
Shimoyama
Tadashi
T
Ohmura
Takeo
T
Sano
Yukari
Y
Nakabayashi
Natsuko
N
Takahashi
Ryoki
R
Fujioka
Toshio
T
Mitchell
Hazel M
HM
Shimoyama
Takashi
T
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D016481:Helicobacter Infections / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology; D044923:Helicobacter hepaticus / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D051381:Rats; D017207:Rats, Sprague-Dawley
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal