Simultaneous detection of Mycoplasma pneumoniae IgG and IgM using dual-label time resolved fluoroimmunoassay

Anti-Mycoplasma pneumoniae (MP) IgM and IgG are useful serological markers for detection of MP infection. In this study, a simultaneous quantification of MP IgM and IgG was performed by time-resolved fluoroimmunoassay (TRFIA). The europium-labeled anti-human IgM and samarium-labeled anti-human IgG were used as tracers, and MP IgM and IgG were recognized in serum samples. After dissociating europium and samarium ions from the immune complex, their fluorescence intensity was recorded and used to calculate the concentrations. The linear range and sensitivity of detection were 2-5500 BU/mL and 0.5 BU/mL for IgM, and 1.5-1500 BU/mL and 0.2 BU/mL for IgG, respectively. The intra- and inter-assay coefficients of variation were 5.14% and 8.41% for IgM, and 5.44% and 8.76% for IgG, respectively. The recovery rate was 94.9-106.8% for IgM and 96.1-109.4% for IgG. The correlation rates of serum detection for 38 respiratory infected patients between dual-label TRFIA and ELISA were 0.9294 and 0.9366 for IgM and IgG, respectively. The coincidence rate between passive particle agglutination and TRFIA is 93.3%. Dual-label TRFIA is a sensitive and reliable technique for measuring MP IgM and IgG levels and could be useful for the early diagnosis of MP infection.