An ELISA for the detection of serum antibodies to Mycoplasma hypopneumoniae in pigs and based on a 43 kDa purified protein derived from the cytoplasmic membrane of M. hyopneumoniae strain J is described. This ELISA (MHPP ELISA) was compared with another recently described (Auspharm ELISA, Sheldrake and Romalis 1992) that is based on column-purified sonicated proteins of strain J. Using sample to negative ELISA ratios of 3 and 4 as cutoffs for inconclusive and positive reactors respectively (compared to 2 and 3 for the Auspharm ELISA), the two tests had high specificity (MHPP 99.6%; Auspharm 100%) in 280 SPF pigs. In 176 pigs from commercial herds with endemic M. hyopneumoniae, the MHPP ELISA showed a higher sensitivity than the Auspharm ELISA in both high lung score (LS > or = 5) (85.5% vs. 69.9%) and low lung score (0 < LS < 5) (57.9% vs. 49%) pigs when the positive cutoff for each test was selected. The sensitivity when the inconclusive cutoff was selected was similar in both tests (85%; 85.7%) when low and high lung score pigs were pooled. Altough the MHPP also gave more positive reactors in 36 pigs from M. hyopneumoniae-infected herds with no lung pathology at slaughter than the Auspharm ELISA (11 vs. 4), the total number of inconclusive and positive reactors in these pigs was similar for both tests (18 vs. 14). The MHPP ELISA gave significantly higher ELISA ratios in infected pigs (up to 17.9) than the Auspharm ELISA (up to 9), and earlier seroconversion in naturally-infected (6-8 weeks vs. 9-10 weeks) and experimentally-infected pigs (2-4 weeks vs. 4-6 weeks post infection).