Despite progress with diagnostic criteria, the type and timing of laboratory tests used to diagnose infective endocarditis (IE) have not been standardized. This is especially true with serological testing. Patients with suspected IE were evaluated by a standard diagnostic protocol. This protocol mandated an evaluation of the patients according to the modified Duke criteria and used a battery of laboratory investigations, including three sets of blood cultures and systematic serological testing for Coxiella burnetii, Bartonella spp., Aspergillus spp., Legionella pneumophila, and rheumatoid factor. In addition, cardiac valvular materials obtained at surgery were subjected to a comprehensive diagnostic evaluation, including PCR aimed at documenting the presence of fastidious organisms. The study included 1,998 suspected cases of IE seen over a 9-year period from

Described here are seven new cases of infective endocarditis due to Escherichia coli, including four involving prosthetic valves, followed by a review of similar cases in the literature. The review identified cases according to the modified Duke's criteria and revealed 16 cases reported before 1960, 5 between 1960 and 1980, and 11 after 1980. Currently, patients diagnosed with E. coli endocarditis are older than the patients diagnosed before 1960 (p<0.05), and they are often diabetic with underlying heart disease. Prosthetic valves are frequently involved (p<0.05), and the principal source of infection is the urinary tract. Surgery is often necessary. The mortality rate associated with this type of infection has decreased since 1960, but it remains high, with 17% calculated for the present series of seven new cases. The data presented here suggest that elderly patients with prior valve disease or prosthetic valve and E. coli urinary tract infection should be examined for endocarditis.

We determined MICs of antibiotics against Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Ehrlichia canis by real-time quantitative PCR. The doubling times of the organisms were established: 19 h for E. chaffeensis, 26 h for A. phagocytophilum, and 28 h for E. canis. In comparison to the reference method for determining sensitivities, which uses Diff-Quick staining, our PCR assay was very sensitive and specific. We confirmed that doxycycline and rifampin are highly active against these bacteria and found variable susceptibilities to fluoroquinolones; A. phagocytophilum was susceptible, but E. canis and E. chaffeensis were only partly susceptible. ␤-Lactam compounds, cotrimoxazole, macrolide compounds, and telithromycin showed no activity against any of the three organisms. Thiamphenicol was found to be more active than chloramphenicol. For the first time, we showed that these three species have numerous point mutations in their 23S RNA genes, with those at positions 754, 2057, 2058, 2059, and 2611 (Escherichia coli numbering) known to confer resistance to macrolide compounds in other bacteria. The role of each of these mutations in resistance to these drugs should be investigated in the future. Our study confirms previous reports that quantitative PCR is a reliable method for determining antibiotic susceptibility; therefore, it might be useful for screening new drugs.

We amplified by PCR and sequenced Streptococcus pneumoniae rpoB from DNA of the cardiac valve of a man who had presented with pneumococcal endocarditis 7 years earlier. Histopathologically, the valve did not show evidence of endocarditis. This case raises the question of persistence of DNA without any evidence of infection.The use of a broad-range bacterial PCR followed by direct sequencing has been successful in the detection of bacterial DNA in excised cardiac tissues of patients with infective endocarditis (IE) (2, 10). 16S rRNA gene amplification by PCR and sequencing were performed on two valves sampled at 7-year intervals in a case of Streptococcus pneumoniae endocarditis. The first valve was obtained after an episode of Pneumococcus IE, and the second was obtained after replacement of the bioprosthetic valve for hemodynamic reasons.Case report. A 48-year-old alcoholic man with a bicuspid aortic valve was admitted in 1994 for endocarditis with meningitis and a cerebral abscess. An aortic vegetation was found, and all three of the blood cultures performed yielded S. pneumoniae. He was treated with ceftriaxone (4 g/day) and thiophenicol, which was replaced with gentamicin (240 mg/day) after 1 week. Worsening aortic insufficiency and persistent fever led to replacement of the aortic valve by pulmonary homograft 15 days later. Macroscopic examination showed a big aortic vegetation (4 cm) with valve destruction and a septal abscess. Microscopic examination showed features typical of IE. Bacterial cultures of the valve were negative.Ceftriaxone (4 g daily) was prescribed for 1 month and then changed to oral amoxicillin (6 g/day) for 1 month. He was monitored as an outpatient with cardiac echography and blood tests 3 and 6 months later and was considered cured. In February 2002, the aortic bioprosthesis was replaced with a mechanical prosthesis because of progressive heart failure. The patient was not febrile, and his white blood cell counts, erythrocyte sedimentation rate, and C-reactive protein levels were normal. Three blood cultures were negative. Macroscopic examination of the valve showed aortic homograft degeneration and calcifications, but no inflammatory cells were found. Gram staining and culture were negative. The two valves were tested at 7-year intervals for bacterial DNA by means of a universal PCR targeting eubacterial 16S rRNA genes. DNA was extracted with Qiagen columns (QIAamp tissue kit; QIAGEN, Hilden, Germany) in accordance with the manufacturer's instructions. PCR amplification with broad-range 16S rRNA gene primers (24) (Table 1), sequencing, and purification of PCR products were performed as previously described (15). After amplification and sequencing of the PCR products, the sequences were compared with those in DNA databases with the BLAST 2.0 program (National Center for Biotechnology Information). The 1995 cardiac valve amplification product showed a nucleotide sequence 100% identical to the 16S rRNA gene sequence of S. pneumoniae (556 of 556 basepairs [bp]; GenBank accession no.AE008...

Scedosporiosis/lomentosporiosis is a devastating emerging fungal infection. Our objective was to describe the clinical pattern and to analyze whether taxonomic grouping of the species involved was supported by differences in terms of clinical presentations or outcomes. We retrospectively studied cases of invasive scedosporiosis in France from 2005 through 2017 based on isolates characterized by polyphasic approach. We recorded 90 cases, mainly related to Scedosporium apiospermum (n = 48), S. boydii/S. ellipsoideum (n = 20), and Lomentospora prolificans (n = 14). One-third of infections were disseminated, with unexpectedly high rates of cerebral (41%) and cardiovascular (31%) involvement. In light of recent Scedosporium taxonomic revisions, we aimed to study the clinical significance of Scedosporium species identification and report for the first time contrasting clinical presentations between infections caused S. apiospermum, which were associated with malignancies and cutaneous involvement in disseminated infections, and infections caused by S. boydii, which were associated with solid organ transplantation, cerebral infections, fungemia, and early death. The clinical presentation of L. prolificans also differed from that of other species, involving more neutropenic patients, breakthrough infections, fungemia, and disseminated infections. Neutropenia, dissemination, and lack of antifungal prescription were all associated with 3-month mortality. Our data support the distinction between S. apiospermum and S. boydii and between L. prolificans and Scedosporium sp. Our results also underline the importance of the workup to assess dissemination, including cardiovascular system and brain. Lay Summary Scedosporiosis/lomentosporiosis is a devastating emerging fungal infection. Our objective was to describe the clinical pattern and to analyze whether taxonomic grouping of the species involved was supported by differences in terms of clinical presentations or outcomes.