[Melioidosis in a Danish tourist returning from North-eastern Thailand].

Melioidosis, an infectious disease caused by Burkholderia pseudomallei, is endemic in South East Asia and Northern Australia. It has a wide clinical diversity, spanning from asymptomatic cases to rapid septic shock and death. We present a case of pulmonary melioidosis in a Danish tourist returning from North-eastern Thailand. The patient was treated with intravenous ceftazidime followed by oral therapy with trimethoprim/sulfamethoxazole and subsequently switched to doxycycline due to abnormal liver function tests and eosinophilia, with no sign of relapse two months after antibiotic cessation.

Combined assay for two-hour identification of Streptococcus pneumoniae and Neisseria meningitidis and concomitant detection of 16S ribosomal DNA in cerebrospinal fluid by real-time PCR.

The main object was to examine the diagnostic performance of a novel combination of a specific real-time PCR (combined real-time PCR) for immediate and simultaneous detection of Streptococcus pneumoniae and Neisseria meningitidis and of a real-time PCR of the 16S rRNA gene (16S DNA). During 12 months, 1015 routine CSF samples were consecutively collected from patients in the County of Aarhus, Denmark. The samples were cultured, examined by microscopy, and, in parallel, CSF DNA was automatically purified and subjected to real-time PCR. Melting curve analysis discriminated between the 2 specific pathogens and 16S DNA positive samples were sequenced. Clinical data were extracted from patients having positive samples. Clinically, 35 of 46 (76%) patients with positive samples had bacterial meningitis. 18 of these 35 patients had a concomitant culture and real-time PCR-positive sample. The remaining 17 patients were either culture positive (n =7) or real-time PCR-positive (n = 10). The aetiology of bacterial meningitis was revealed by microscopy in 18/35 (51.4%), culture in 24/35 (68.6%) and combined real-time PCR in 27/35 (77.1%) patients, respectively. In conclusion, the combined real-time PCR strategy is superior to microscopy and a valuable supplement to routine culture to establish the aetiology of bacterial meningitis.

Diagnosis of ventricular drainage-related bacterial meningitis by broad-range real-time polymerase chain reaction.

OBJECTIVE: To compare a broad-range real-time polymerase chain reaction (PCR) diagnostic strategy with culture to evaluate additional effects on the etiological diagnosis and the quantification of the bacterial load during the course of ventricular drainage-related bacterial meningitis (VR-BM). METHODS: We applied a PCR that targeted conserved regions of the 16S ribosomal ribonucleic acid gene to cerebrospinal fluid (CSF) samples from patients with external ventricular drainage or a ventriculoperitoneal shunt during the course of VR-BM. We compared the PCR results with CSF cultures. A total of 350 routine CSF samples were consecutively collected from 86 patients. The CSF deoxyribonucleic acid was automatically purified and subjected to PCR. Amplicons from the PCR samples that were positive for VR-BM were subsequently deoxyribonucleic acid sequenced for final identification. Clinical data were extracted from patient files. RESULTS: Sixteen patients had at least one VR-BM-positive sample as diagnosed from culture or PCR. Nineteen episodes were diagnosed with signs of VR-BM (n = 16 patients) or were determined to be contaminated (n = 3 patients). Four episodes of VR-BM were diagnosed via PCR alone and were predominantly caused by gram-negative pathogens, five episodes were diagnosed via culture alone, and seven episodes were diagnosed via both culture and PCR. Five patients had mixed infections. Overall, 71 samples were positive for VR-BM as indicated by either one or both of the methods. Eighteen CSF samples were VR-BM positive as indicated by culture alone, and 21 CSF samples were positive as indicated via PCR alone. CONCLUSIONS: Culture supplemented with broad-range, real-time PCR may increase the number of etiologically diagnosed VR-BM episodes, particularly when these are caused by gram-negative bacteria.

Broad-range real time PCR and DNA sequencing for the diagnosis of bacterial meningitis.

Rapid aetiological diagnosis of bacterial meningitis is crucial for the early targeting of antimicrobial and adjuvant therapy. Broad-range polymerase chain reaction (PCR) targeting the 16S rRNA gene allows aetiological diagnosis of bacterial meningitis when applied to cerebrospinal fluid (CSF). We assessed the additional diagnostic effect of applying a novel broad-range real time PCR and subsequent DNA sequencing to culture, microscopy, and broad-range conventional PCR on CSF in patients with suspected bacterial meningitis. Broad-range conventional PCR and broad-range real time PCR with subsequent DNA sequencing were applied to 206 CSF specimens collected consecutively from 203 patients aged 6 d to 86 y. Patients' charts were reviewed for clinical information. 17 pathogens were identified by PCR and DNA sequencing or culture. Three specimens were negative by culture but positive by broad-range real time PCR. Three specimens were positive by culture but negative by broad-range real time PCR. Compared with culture, the sensitivity of broad-range real time PCR was 86%, and the specificity 98%. Conventional PCR resulted in a sensitivity of 64% and specificity of 98%. Broad-range real time PCR was generally comparable to culture of CSF and may be a useful supplement, particularly when antimicrobial therapy has been administered. Broad-range real time PCR was more sensitive than broad-range conventional PCR and microscopy.

Crowding as a risk factor of meningococcal disease in Danish preschool children: a nationwide population-based case-control study.

Meningococcal disease (MD) remains an important health problem. Crowding has been suggested to be a risk factor for MD in children, but the evidence is relatively sparse. We performed a nationwide nested case-control study comprising 1222 children with MD and 24,549 population controls. We identified MD cases younger than 6 y in the Danish National Hospital Discharge Registry from 1980 to 1999, and obtained information on household density as a measure of crowding, per capita income and other potential confounders through The Danish Civil Registration System and social registries. The risk of MD associated with household density was estimated by conditional logistic regression for children less than 1 y of age (infants) and children aged 1 to 5 y, respectively. The risk of MD increased with increasing household density. In both age groups, the crude OR was 1.8 (95% confidence interval [CI]: 1.4-2.3) at a density of less than 20 m2 per person compared with the reference of more than 50 m2 per person. The adjusted OR for MD was 1.5 (95% CI: 1.1-1.9) for infants, and 1.5 (95% CI: 1.1-2.0) for children older than 1 y. Household density appears to be a risk factor of MD in preschool children.

Prospective evaluation of the quality of oral anticoagulation management in an outpatient clinic and in general practices.

OBJECTIVE: To compare the quality of oral anticoagulant therapy (QOAT), before and after referral of patients on oral anticoagulant therapy (OAT) from a hospital outpatient clinic (HOC) to general practitioners (GPs). DESIGN: Prospective observational study. Patients were identified by using the Laboratory Information System (LIS), containing all prescribed International Normalised Ratio of Prothrombin Time (INR) tests, from the HOC and GPs in the hospital submission area. SETTING: The HOC in a rural hospital, Aarhus County, Denmark (55,000 inhabitants), and GPs in the submission area. SUBJECTS: 124 OAT patients (59.7% males. Median age 70.0: 25-75 percentile: 62.0-76.0). MAIN OUTCOME MEASURE: The QOAT in terms of time spent within therapeutic INR interval (TI). The QOAT was compared 8 months before with 8 months after altering the monitoring organization. For patients monitored less than 8 months before the alteration, the QOAT was compared to a corresponding time period after the alteration. RESULTS: We identified 124 OAT patients, and found a significant increase in the QOAT from 65.0% before to 69.1% after referral of the patients to the GPs (P<.0001). In 75 patients with full follow-up, the QOAT increased from 67.5% before to 69.7% after the alteration (P<.0001). CONCLUSION: The results indicate that the QOAT in this geographical area is adequate, and that the quality performed by the GPs was at least as good as in the HOC. In order to document and increase the QOAT, continuous quality surveillance using the LIS has been initiated.