16S rDNA PCRs were performed once a week at the request of clinicians or by microbiologists’ initiative and systematically on cardiac valve samples with suspected endocarditis when the sample’s cultures were negative. Briefly, purified DNA was extracted from clinical samples using MagNA Pure (Roche), PCRs were performed with the LightCycler® 2.0 (Roche) instrument with 27F/16S1RRB primers and sequencing reactions were performed using the Sanger method on an ABI 3730 XL system (Applied Biosystems) as described in detail [23]. A microbiologist analyzed the results using the online tool leBIBIQBPP [24] (https://umr5558-bibiserv.univ-lyon1.fr/lebibi/lebibi.cgi) to compare the tested sequence to sequences derived from the International Nucleotide Sequence Database Collaboration (http://www.insdc.org) – including the GenBank® database and EMBL-ENI –, allowing construction of phylogenetic unrooted approximate maximum-likelihood trees and bacterial final identification by using the minimal patristic distance and the nearest reference strain in the tree as the decision criteria. Conclusions were transmitted to the clinicians.

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