Serological diagnosis of enzootic pneumonia of swine by a double-sandwich enzyme-linked immunosorbent assay using a monoclonal antibody and recombinant antigen (P46) of Mycoplasma hyopneumoniae

• Published in: Veterinary microbiology Vol. 105; no. 3; pp. 251 - 259
• Main Authors: Okada, Munenori, Asai, Tetsuo, Futo, Satoshi, Mori, Yasuyuki, Mukai, Tetsuya, Yazawa, Shigeto, Uto, Takehiko, Shibata, Isao, Sato, Shizuo
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 25-02-2005; Elsevier Science
• Subjects: Animals; Antibodies, Bacterial - biosynthesis; Antibodies, Bacterial - blood; Antibodies, Monoclonal; antibody detection; Antigens, Bacterial - genetics; Antigens, Bacterial - immunology; Applied microbiology; Bacteria; Bacterial Proteins - genetics; Bacterial Proteins - immunology; Bacteriology; Biological and medical sciences; blood serum; Complement Fixation Tests - methods; Complement Fixation Tests - veterinary; Cross Reactions; disease detection; disease diagnosis; ELISA; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; Escherichia coli; Female; field experimentation; Fundamental and applied biological sciences. Psychology; Immunoblotting - veterinary; immunologic techniques; Male; methodology; Microbiology; Miscellaneous; monoclonal antibodies; Monoclonal antibody; Mycoplasma hyopneumoniae; Mycoplasma hyopneumoniae - immunology; Mycoplasma hyopneumoniae - isolation & purification; Mycoplasma pneumoniae; Parity; Pig; Pneumonia of Swine, Mycoplasmal - diagnosis; Pneumonia of Swine, Mycoplasmal - epidemiology; Recombinant antigen; recombinant antigens; Recombinant Proteins - immunology; Sensitivity and Specificity; serodiagnosis; Seroepidemiologic Studies; Specific Pathogen-Free Organisms; Swine; swine diseases; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects); Bacteria; Monoclonal antibody; Mycoplasma hyopneumoniae; Recombinant antigen; ELISA; Pig; Lung disease; Pneumonia; Microbiology; Respiratory disease; Mycoplasmataceae; Mollicutes; Mycoplasmatales; Vertebrata; Mammalia; Swine; Artiodactyla; Diagnosis; Recombinant protein; Veterinary; ELISA assay; Ungulata
• ISSN: 0378-1135; 1873-2542
• DOI: 10.1016/j.vetmic.2004.11.006

To facilitate the control of enzootic pneumonia (EP) of swine caused by
Mycoplasma hyopneumoniae, the complement fixation (CF) test has been used for the detection of
M. hyopneumoniae antibodies. However, the CF test is a cumbersome and time-consuming technique and cross-reactivity are major drawbacks associated with this method. To circumvent these drawbacks, we have developed a double-sandwich enzyme-linked immunosorbent assay (ELISA), consisting of purified monoclonal antibody (Mab) against the 46 kDa surface antigen (P46) of
M. hyopneumoniae and recombinant P46 protein expressed in
Escherichia coli, for the detection of antibodies to
M. hyopneumoniae in serum samples from pigs experimentally inoculated with
M. hyopneumoniae and from naturally infected pigs, and compared the practical usefulness of ELISA using the CF test. In experimentally inoculated pigs, the CF and ELISA antibodies were detected at almost the same time, and a good correlation was demonstrated between the CF test and the ELISA. In a survey conducted on field samples, the seropositivity by ELISA in pigs of age 2–6 months was increased. At the time of slaughter, approximately 80% of the animals were seropositive for ELISA. However, a gradual decrease in the prevalence of ELISA positive samples was observed in sows with increasing parity. No correlation was seen between the results obtained with the two methods in the clinical samples. The CF test appears to have limited value for the diagnosis of EP in conventional herds because nonspecific reactions were frequently observed. Therefore, this ELISA is a useful alternative to the CF test currently used for the diagnosis of EP.