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Journal of Clinical Microbiology, March 2003, p. 919-925, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.919-925.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Diagnosis of Bartonella Endocarditis by a Real-Time Nested PCR Assay Using Serum

Zaher Zeaiter, Pierre-Edouard Fournier, Gilbert Greub, and Didier Raoult*

Unité des Rickettsies, IFR 48, CNRS UMR 6020, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 5, France

Received 20 June 2002/ Returned for modification 20 August 2002/ Accepted 2 December 2002

Bartonella endocarditis is a severe disease for which blood cultures frequently remain negative. We tested three PCR assays by using specimens of serum sampled early during the disease from 43 patients diagnosed in our laboratory as having Bartonella endocarditis on the basis of serological, culture, and/or valvular molecular detection. We tested a two-step nested PCR (TSN-PCR), a one-step nested PCR (OSN-PCR) with a regular thermal cycler, and a one-step nested PCR with the LightCycler (LCN-PCR). These assays were performed with primers derived from the riboflavin synthase-encoding gene ribC, never before amplified in our laboratory. Due to contamination of negative controls, the results of the TSN-PCR were not interpretable, and this technique was no longer considered. The LCN-PCR had a specificity of 100% and a sensitivity of 58.1%, higher than those of the OSN-PCR (18.6%; P < 0.01) and prolonged blood culturing (7.1%; P < 0.01). The LCN-PCR results correlated strictly with those of other direct diagnostic tests, when available, and identified the causative species for six patients previously diagnosed on the basis of serological analysis only. The efficacy of the LCN-PCR was not influenced by antibiotics (P = 0.96) but was altered by prolonged storage of serum specimens at -20°C (P = 0.04). Overall, the LCN-PCR is specific and more sensitive than traditional methods (i.e., culturing and/or PCR with EDTA-treated blood). It can easily be applied to the diagnosis of patients with suspected Bartonella endocarditis, especially when only serum is available.

* Corresponding author. Mailing address: Unité des Rickettsies, IFR 48, CNRS UMR 6020, Faculté de Médecine, Université de la Méditerranée, 27 Blvd. Jean Moulin, 13385 Marseille Cedex 5, France. Phone: (33) 491 38 55 17. Fax: (33) 491 83 03 90. E-mail: Didier.Raoult{at}medecine.univ-mrs.fr.

Journal of Clinical Microbiology, March 2003, p. 919-925, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.919-925.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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