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Clinical and Vaccine Immunology, March 2008, p. 447-451, Vol. 15, No. 3
1071-412X/08/$08.00+0     doi:10.1128/CVI.00379-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Production of Recombinant Antigens of Ureaplasma parvum Serotypes 3 and 6 for Development of a Serological Assay{triangledown}

E. Vancutsem,* F. Echahidi, K. Van Geel, G. Muyldermans,{dagger} O. Soetens, and A. Naessens

Department of Microbiology, Universitair Ziekenhuis Brussel, Laarbeeklaan 101, 1090 Jette, Belgium

Received 13 September 2007/ Returned for modification 10 October 2007/ Accepted 5 December 2007

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 by PCR and cloned in a pTrcHis TOPO plasmid. Purified recombinant proteins were evaluated in Western blotting and enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies and human sera. Our approach was successful in the production of the recombinant MBAs (rMBAs) for serotypes 3 and 6. The antigens tested positive with serotype-specific monoclonal antibodies in Western blotting and in ELISA. Prominent reactions were detected with the rMBAs and their homologous monoclonal antibodies. Strong cross-reactions were visible in ELISA between rMBA 3 and the monoclonal antibodies from the other U. parvum serotypes. A weak cross-reaction was seen with rMBA 3 and the monoclonal antibody from serotype 4. rMBA 6 showed cross-reaction only 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 three (15%) of the sera from culture-negative women reacted with the rMBA. The positive reactions were observed only with rMBA 6. These preliminary tests showed the potential usefulness of the rMBAs produced for detecting an antibody response against Ureaplasma antigens.

* Corresponding author. Mailing address: Department of Microbiology, Universitair Ziekenhuis Brussel, Laarbeeklaan 101, 1090 Jette, Belgium. Phone: 32(0)24775003. Fax: 32(0)24775015. E-mail: Ellen.Vancutsem{at}uzbrussel.be

{triangledown} Published ahead of print on 19 December 2007.

{dagger} Present address: Virco BVBA, Generaal De Wittelaan L11 B4, 2800 Mechelen, Belgium.

Clinical and Vaccine Immunology, March 2008, p. 447-451, Vol. 15, No. 3
1071-412X/08/$08.00+0     doi:10.1128/CVI.00379-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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