1. Differential recognition of the multiple banded antigen isoforms across Ureaplasma parvum and Ureaplasma urealyticum species by monoclonal antibodies.
- Author
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Aboklaish AF, Ahmed S, McAllister D, Cassell G, Zheng XT, and Spiller OB
- Subjects
- Antibodies, Bacterial isolation & purification, Antibodies, Monoclonal isolation & purification, Antigens, Bacterial genetics, Bacterial Proteins genetics, DNA, Bacterial, Escherichia coli genetics, Humans, Immunoblotting, Polymerase Chain Reaction, Protein Isoforms immunology, Sequence Analysis, DNA, Serogroup, Ureaplasma Infections diagnosis, Ureaplasma Infections immunology, Ureaplasma Infections microbiology, Antibodies, Bacterial immunology, Antibodies, Monoclonal immunology, Antigens, Bacterial immunology, Immunologic Techniques, Ureaplasma immunology, Ureaplasma urealyticum immunology
- Abstract
Two separate species of Ureaplasma have been identified that infect humans: Ureaplasma parvum and Ureaplasma urealyticum. Most notably, these bacteria lack a cell wall and are the leading infectious organism associated with infection-related induction of preterm birth. Fourteen separate representative prototype bacterial strains, called serovars, are largely differentiated by the sequence of repeating units in the C-terminus of the major surface protein: multiple-banded antigen (MBA). Monoclonal antibodies that recognise single or small groups of serovars have been previously reported, but these reagents remain sequestered in individual research laboratories. Here we characterise a panel of commercially available monoclonal antibodies raised against the MBA and describe the first monoclonal antibody that cross-reacts by immunoblot with all serovars of U. parvum and U. urealyticum species. We also describe a recombinant MBA expressed by Escherichia coli which facilitated further characterisation by immunoblot and demonstrate immunohistochemistry of paraffin-embedded antigens. Immunoblot reactivity was validated against well characterised previously published monoclonal antibodies and individual commercial antibodies were found to recognise all U. parvum strains, only serovars 3 and 14 or only serovars 1 and 6, or all strains belonging to U. parvum and U. urealyticum. MBA mass was highly variable between strains, consistent with variation in the number of C-terminal repeats between strains. Antibody characterisation will enable future investigations to correlate severity of pathogenicity to MBA isoform number or mass, in addition to development of antibody-based diagnostics that will detect infection by all Ureaplasma species or alternately be able to differentiate between U. parvum, U. urealyticum or mixed infections., (Copyright © 2016 Elsevier B.V. All rights reserved.)
- Published
- 2016
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