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Journal of Clinical Microbiology, February 2003, p. 763-766, Vol. 41, No. 2
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.2.763-766.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Diagnosis of Infective Endocarditis by PCR Amplification and Direct Sequencing of DNA from Valve Tissue

Valérie Gauduchon,1 Lara Chalabreysse,2 Jerome Etienne,1 Marie Célard,1 Yvonne Benito,1 Hubert Lepidi,3 Françoise Thivolet-Béjui,2 and François Vandenesch1*

Laboratoire de Bactériologie,1 Service d'Anatomie et de Cytologie Pathologiques, Hôpital Louis Pradel, 69394 Lyon Cedex 03,2 Laboratoire d'Histologie and Unité des Rickettsies, CNRS UMR 6020, Faculté de Médecine, 13385 Marseille Cedex 5, France3

Received 1 August 2002/ Returned for modification 30 September 2002/ Accepted 5 November 2002

We used broad-range eubacterial PCR amplification followed by direct sequencing to identify microbial pathogens in heart valve material from 29 patients with histologically confirmed infective endocarditis and 23 patients free of infective endocarditis. Microorganisms cultured by conventional techniques matched those identified by PCR in 21 cases. PCR alone identified the causative agent in three cases (Streptococcus bovis, Staphylococcus cohnii, and Coxiella burnetii), allowing better patient management. PCR corrected the initial bacteriological diagnosis in three cases (Streptococcus bovis, Streptococcus mutans, and Bartonella henselae). Among the 29 cases of histologically confirmed infective endocarditis, PCR findings were positive in 27 cases and were consistent with the bacterial morphology seen at Gram staining (26 cases) or with the results obtained by immunohistologic analysis with an anti-C. burnetii monoclonal antibody (one case). In two other cases of histologically confirmed infective endocarditis, PCR remained negative in a blood culture-negative case for which no bacteria were seen at histological analysis and in one case with visualization of cocci and blood cultures positive for Enterococcus faecalis. Ten clinical diagnoses of possible infective endocarditis were ruled out by histopathological analysis of the valves and subsequently by PCR. PCR was negative in 13 of the 14 patients in whom infective endocarditis was rejected on clinical grounds; the other patient was found to have Coxiella burnetii infective endocarditis on the basis of PCR and histopathological analysis and was subsequently included in the group of 29 definite cases. In total, PCR contributed to the diagnosis and management of infective endocarditis in 6 of 29 (20%) cases.

* Corresponding author. Mailing address: Laboratoire de Bactériologie, Faculté de Médecine, Rue Guillaume Paradin, 69372 Lyon Cedex 08, France. Phone: (33) 47 877 86 57. Fax: (33) 47 877 86 58. E-mail: denesch{at}univ-lyon1.fr.

Journal of Clinical Microbiology, February 2003, p. 763-766, Vol. 41, No. 2
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.2.763-766.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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