Development of an ELISA technique for serodiagnosis of bovine paratuberculosis.
A local clinical Mycobacterium avium ssp. paratuberculosis (M. paratuberculosis) isolate was cultivated in large-scale culture. A procedure for efficient preparation of a lipoarabinomannan-containing antigen was developed and standardized; 25 mg of purified antigen were obtained per gram of bacterial wet weight. An ELISA based on this antigen was developed. Intra- and interassay variation were determined to be 20% and 27%, respectively. The ELISA was evaluated using the sera of groups of 39 non-randomly selected and 92 randomly selected animals from which ileocaecal lymph nodes were cultured to isolate viable M. paratuberculosis. Combining the results of both groups a positive predictive value of 74% and a negative predictive value of 99% were calculated. The ELISA was licensed by the German regulatory agencies and, in a direct comparison, was found to be superior to both other licensed assays.
Jark
U
U
Institut für Mikrobiologie und Tierseuchen, Tierärztliche Hochschule Hannover, Germany.
Ringena
I
I
Franz
B
B
Gerlach
G F
GF
Beyerbach
M
M
Franz
B
B
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology; D015153:Blotting, Western; D002417:Cattle; D003168:Complement Fixation Tests / Q000379:methods / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005243:Feces / Q000382:microbiology; D008070:Lipopolysaccharides / Q000276:immunology; D008198:Lymph Nodes / Q000382:microbiology; D016927:Mycobacterium avium subsp. paratuberculosis / Q000276:immunology / Q000302:isolation & purification; D010283:Paratuberculosis / Q000175:diagnosis* / Q000473:pathology; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D008070:Lipopolysaccharides; C050016:lipoarabinomannan