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Use of the P167 recombinant antigen for serodiagnosis of Helicobacter bilis
Lon V Kendall1, Sunlian Feng, Emir Hodzic
1Center for Comparative Medicine, Schools of Medicine and Veterinary Medicine, University of California, Davis, Davis, California 95616, USA.
Abstract:
Helicobacter bilis is widespread among research mouse colonies. Serodiagnosis of Helicobacter infections involves use of bacterial lysates or membrane antigen preparations that lack specificity, necessitating the need to identify a specific and sensitive antigen. A previously reported recombinant protein (P167) was evaluated for use as an H. bilis-specific antigen for serologic testing. Seventy-six mice naturally infected with Helicobacter spp. were identified from commercially bred or sentinel mice. Infection was confirmed and speciated by use of cecal specimen culture and fecal polymerase chain reaction (PCR) analysis, followed by restriction enzyme digest of the amplicon. Forty-one mice were determined to be monoinfected with H. bilis, 27 mice were determined to be monoinfected with H. hepaticus, and eight mice were infected with another species of Helicobacter. Serum was diluted 1:100 to evaluate the immunoreactivity to enzyme-linked immunosorbent assay preparations of H. bilis membrane extract and the immunodominant C and D fragments of the p167 gene. The sensitivity was greatest for the membrane extract preparation (76%), whereas sensitivity to the P167C and D recombinants was lower (62 and 51%, respectively). However, the specificity of the membrane extract preparation was low (87%), compared with the much improved specificity of the recombinant P167C and D fragments (96 and 96%, respectively). These findings suggest that the recombinant P167C and D fragments of the p167 gene product from H. bilis can be used as specific reagents in the serodiagnosis of H. bilis infection in mice.
Insights
Specific recombinant proteins P167C and P167D offer improved specificity for diagnosing Helicobacter bilis infections in mice, addressing limitations of current serodiagnostic methods.
Area of Science:
- Veterinary Microbiology
- Immunology
- Animal Health Diagnostics
Background:
- Helicobacter bilis is a common pathogen in research mouse colonies.
- Current serodiagnostic methods using bacterial lysates or membrane antigens lack specificity.
- There is a need for a specific and sensitive antigen for accurate H. bilis detection.
Purpose of the Study:
- To evaluate a previously reported recombinant protein (P167) and its fragments (P167C and P167D) as specific antigens for H. bilis serodiagnosis.
- To compare the sensitivity and specificity of these recombinant antigens against H. bilis membrane extract.
- To determine the utility of P167 fragments in identifying H. bilis infections in mice.
Main Methods:
- Seventy-six infected mice were identified and infections confirmed/speciated using cecal culture and fecal PCR.
- Serum samples were tested via enzyme-linked immunosorbent assay (ELISA) against H. bilis membrane extract and P167C/P167D recombinant proteins.
- Sensitivity and specificity of each antigen preparation were calculated.
Main Results:
- H. bilis membrane extract showed the highest sensitivity (76%) but low specificity (87%).
- Recombinant P167C and P167D fragments demonstrated lower sensitivity (62% and 51%) but significantly higher specificity (96% for both).
- The P167C and P167D fragments were highly specific for detecting H. bilis infections.
Conclusions:
- Recombinant P167C and P167D fragments are specific reagents for serologic diagnosis of H. bilis infection in mice.
- These recombinant antigens overcome the specificity limitations of traditional membrane antigen preparations.
- The findings support the use of P167C and P167D for improved accuracy in mouse Helicobacter diagnostics.
