Production of recombinant antigens of Ureaplasma parvum serotype 3 and 6 for the development of a serological assay

Universitair Ziekenhuis Brussel, Department: Microbiology, Laarbeeklaan 101, 1090 Jette (Belgium) and VIRCO BVBA, Generaal De Wittelaan L11 B4, 2800 Mechelen (Belgium)

^* To whom correspondence should be addressed. Email: Ellen.Vancutsem{at}uzbrussel.be.

	Abstract

Recombinant antigens of Ureaplasma parvum serotypes 3 and 6 were produced in order to develop a serological assay for Ureaplasma antibody detection. The genes of the multiple banded antigen (MBA) were amplified with PCR and cloned in a pTrcHis TOPO plasmid. Purified recombinant proteins were evaluated in Western blot and ELISA with monoclonal antibodies and human sera.

Our approach was successful in the production of the recombinant MBA (rMBA) for serotypes 3 and 6. The antigens tested positive with serotype specific monoclonal antibodies in Western blot and in ELISA. Prominent reactions were detected with the recombinant multiple banded antigen (rMBA) and their homologous monoclonal antibodies. Strong cross reactions were visible in ELISA between the rMBA 3 and the monoclonal antibodies from the other U. parvum serotypes. A weak cross reaction was seen with the rMBA 3 and the monoclonal antibody from serotype 4. The rMBA 6 showed only cross reaction with the monoclonal antibody from U. parvum serotype 1. Fifty one percent of the sera obtained from culture positive women, reacted with one or both rMBAs. Only 3 (15%) of the sera from culture negative women reacted with the rMBA. These positive reactions were only observed with rMBA 6. These preliminary tests showed the potential usefulness of the produced rMBAs for detecting an antibody response against Ureaplasma antigens.