Detection and screening of Salmonella enteritidis-infected chickens with recombinant flagellin.
Screening and identification of Salmonella enteritidis in commercial poultry flocks have assumed principal roles in preventing transmission of this pathogen to humans from hen eggs. Serologic diagnosis of S. enteritidis infection in commercial flocks currently relies on laboratory-based tests for detection of antibodies to the lipopolysaccharide, whole flagella, and bacteria. We amplified a sequence from the g,m flagellin of S. enteritidis, followed by cloning, expression, and purification of the protein. The recombinant protein was first characterized by western blot and subsequently evaluated as enzyme-linked immunosorbent assay (ELISA) antigen for detection of S. enteritidis infection. A total number of 49 positive sera and 40 negative sera were tested for ELISA validation. A cutoff value of 0.14 was shown to be sufficient to discriminate the negative and positive sera. Results obtained by testing sera raised against different bacterial strains/serotypes further confirmed that this recombinant flagellin-based ELISA was indeed specific for the detection of S. enteritidis. Both sensitivity and specificity of the developed ELISA test were comparable with a commercially available test, indicating that it is a highly promising and reliable diagnostic tool for S. enteritidis infection.
Yap
L F
LF
Animal Health Biotechnology, Institute of Molecular Agrobiology, National University of Singapore, Singapore.
Low
S
S
Liu
W
W
Loh
H
H
Teo
T P
TP
Kwang
J
J
D000818:Animals; D000942:Antigens, Bacterial / Q000097:blood; D002645:Chickens; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005260:Female; D005408:Flagellin / Q000235:genetics / Q000276:immunology*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology; D011994:Recombinant Proteins / Q000276:immunology; D015203:Reproducibility of Results; D012481:Salmonella Infections, Animal / Q000175:diagnosis* / Q000276:immunology; D012477:Salmonella enteritidis / Q000276:immunology*; D012680:Sensitivity and Specificity; D012698:Serologic Tests
D000942:Antigens, Bacterial; D011994:Recombinant Proteins; D005408:Flagellin