Development and validation of a SIgA-ELISA for the detection of Mycoplasma hyopneumoniae infection.

An alternative indirect enzyme-linked immunosorbent assay (ELISA) for the detection of Mycoplasma hyopneumoniae secretory IgA (SIgA) antibody (SIgA-ELISA) was developed using an adhesin (P97R1) of M. hyopneumoniae produced in Escherichia coli. The SIgA-ELISA assay was validated by the comparison with a nested-PCR assay and a commercial M. hyopneumoniae antibody detection kit (IgG-ELISA). Two hundred and sixty nasal swab samples, bronchoalveolar lavage fluids or serum samples were prepared for SIgA-ELISA validation from a M. hyopneumoniae-free farm, a M. hyopneumoniae vaccinated farm and two M. hyopneumoniae contaminated farms. The results showed that the SIgA-ELISA assay could distinguish the M. hyopneumoniae infection from M. hyopneumoniae vaccinated pigs, which was impossible for the current commercial M. hyopneumoniae antibody detection kits. The diagnostic sensitivity (DSN), specificity (DSP) and accuracy of the SIgA-ELISA were 97.0%, 94.4% and 95.8%, respectively and were compared with nested-PCR on 260 field nasal swab samples. The results of repeatability tests revealed that the coefficients of variation of swab samples within and between runs were less than 10%. This SIgA-ELISA is a needle-free detection methodology for large-scale surveys of M. hyopneumoniae infection. (c) 2009 Elsevier B.V. All rights reserved.