PURPOSE: This study aimed to improve the serological identification of urogenital mycoplasmas through the development of MUREAPLEXE, a recombinant protein-based multiplex ELISA allowing the simultaneous and specific detection of antibodies against Metamycoplasma hominis, Ureaplasma parvum and Ureaplasma urealyticum. METHODS: Genes encoding the major surface antigens Vaa and P120' of M. hominis, MBA3 of U. parvum and MBA8 of U. urealyticum were cloned into the prokaryotic expression vector pGEX-4T-1 and expressed in E. coli BL21. Recombinant GST-fusion proteins were purified by affinity chromatography using Glutathione-Sepharose 4B beads and were used to optimize an indirect multiplex ELISA. Both assays were subsequently evaluated using 162 patient sera that had been previously characterized by indirect immunofluorescence (IIF) and Western blot analysis. RESULTS: Relative to IIF, both ELISA assays demonstrated high sensitivity. All tested sera showed reactivity with their corresponding recombinant antigens. Statistical analyses demonstrated a significant correlation between the two ELISA tests. However, MUREAPLEXE demonstrated markedly higher specificity. CONCLUSION: Given its high sensitivity and specificity for the simultaneous serological identification of M. hominis, U. parvum, and U. urealyticum, MUREAPLEXE may provide a robust and reliable complementary tool for the monitoring of urogenital mycoplasmas.
Abassi et al. (Tue,) studied this question.