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Clinical and Diagnostic Laboratory Immunology, July 2000, p. 617-624, Vol. 7, No. 4
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Differentiation of Bartonella Species by a Microimmunofluorescence Assay, Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis, and Western Immunoblotting

Zhongxing Liang and Didier Raoult*

Unité des Rickettsies, CNRS UPRES-A 6020, Faculté de Médecine, Université de la Mediterranée, 13385 Marseille Cédex, France

Received 3 December 1999/Returned for modification 4 April 2000/Accepted 8 May 2000

Bartonella species can be differentiated by microimmunofluorescence assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotting with murine polyclonal antisera to Bartonella henselae, B. quintana, B. elizabethae, and B. bacilliformis. A pairwise comparison on the basis of SDS-PAGE protein profiles demonstrated similarity values for proteins of different Bartonella species ranging from 28.6 to 86.4%. Antigenic relationships revealed by immunoblotting with murine antisera were equivalent to those of proteins observed by SDS-PAGE. A dendrogram obtained on the basis of protein bands of SDS-polyacrylamide gels showed that Bartonella species could be divided into three groups. B. bacilliformis was distinct from all other Bartonella species; B. grahamii, B. taylorii, B. doshiae, and B. vinsonii formed a cluster, as did B. henselae, B. quintana, B. elizabethae, and B. clarridgeiae. These relationships were consistent with those revealed by parsimony trees derived from 16S rRNA and gltA gene sequencing. SDS-PAGE analysis showed that 120-, 104-, 85-, 71-, 54-, 47-, 40-, 33-, 30-, and 19-kDa proteins were present in all species, with the 54-kDa protein being the most dominant. Proteins with a molecular mass of less than 54 kDa allow the differentiation of species and are a possible target for future species-specific antibodies and antigens.

* Corresponding author. Mailing address: Unité des Rickettsies, Faculté de Médecine, 27 Blvd. Jean Moulin, 13385 Marseille Cedex 5, France. Phone: (33) 4 91 32 43 75. Fax: (33) 4 91 83 03 90. E-mail: Didier.Raoult{at}medecine.univ-mrs.fr.

Clinical and Diagnostic Laboratory Immunology, July 2000, p. 617-624, Vol. 7, No. 4
1071-412X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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