Case report: tick-borne encephalitis in two Dutch travellers returning from Austria, Netherlands, July and August 2011.

Tick-borne encephalitis (TBE) is not endemic in the Netherlands and diagnostics are seldom requested. Here, we report about the rare event of TBE in two Dutch travellers returning from Austria in July and August 2011. This report serves to create awareness among physicians to consider travel-related TBE in their differential diagnosis of patients with neurological disease returning from TBE virus endemic regions and to promote awareness among professionals advising travellers.

Prevalence, characterisation and clinical profiles of Shiga toxin-producing Escherichia coli in The Netherlands.

Detection of Shiga toxin-producing Escherichia coli (STEC) in The Netherlands is traditionally limited to serogroup O157. To assess the relative importance of STEC, including non-O157 serogroups, stool samples submitted nationwide for investigation of enteric pathogens or diarrhoea were screened with real-time PCR for the presence of the Shiga toxin genes. Patients were selected if their stool contained blood upon macroscopic examination, if they had a history of bloody diarrhoea, were diagnosed with haemolytic uraemic syndrome, or were aged <6 years (irrespective of the bloody aspect of the stool). PCR-positive stools were forwarded to a central laboratory for STEC isolation and typing. In total, 4069 stools were examined, with 68 (1.7%) positive PCR results. The highest prevalence was for stools containing macroscopic blood (3.5%), followed by stools from patients with a history of bloody diarrhoea (2.4%). Among young children, the prevalence (1.0%) was not significantly higher than among random, non-bloody, stool samples from diarrhoeal patients (1.4%). STEC strains were isolated from 25 (38%) PCR-positive stools. Eleven O-serogroups were detected, including five STEC O157 strains. As serogroup O157 represented only 20% of the STEC isolates, laboratories should be encouraged to use techniques enabling them to detect non-O157 serogroups, in parallel with culture, for isolation and subsequent characterisation of STEC strains for public health surveillance and detection of outbreaks.

The 13carbon urea breath test for the diagnosis of Helicobacter pylori infection in subjects with atrophic gastritis: evaluation in a primary care setting.

BACKGROUND: (13)Carbon urea breath testing is reliable to detect current infection with Helicobacter pylori but has been reported to be of limited value in selected patients with atrophic body gastritis or acid-lowering medication. AIM: To evaluate the accuracy of (13)carbon urea breath testing for H. pylori detection in 20 asymptomatic patients with histologically confirmed atrophic body gastritis in a primary care setting. METHODS: (13)Carbon urea breath testing and serology were compared with H. pylori culture of a corpus biopsy as reference test. RESULTS: All tests were in agreement in 12 patients, being all positive in six and all negative in six. One patient was positive for serology and culture but negative for (13)carbon urea breath testing, five patients had only positive serology and two patients had only positive (13)carbon urea breath testing. (13)Carbon urea breath testing showed an accuracy with culture of 85% and anti-H. pylori serology with culture of 75%. (13)Carbon urea breath testing carried out in patients with positive serology showed an accuracy of 92%. Receiver operating characteristic curve analysis of (13)carbon urea breath testing shows optimal discrimination at the prescribed cut-off value. CONCLUSIONS: (13)Carbon urea breath testing can be used as diagnostic H. pylori test in asymptomatic patients with atrophic body gastritis, preferably in addition to serology, to select subjects for anti-H. pylori therapy.

Increasing resistance to fluoroquinolones in escherichia coli from urinary tract infections in the netherlands.

In continuous surveillance of routine samples from five Dutch laboratories, we studied resistance to the antibiotics most commonly prescribed for urinary tract infections (UTI) in The Netherlands, namely norfloxacin, amoxycillin, trimethoprim and nitrofurantoin, from 1989 to 1998 in >90000 Escherichia coli isolates. Resistance to norfloxacin increased from 1.3% in 1989 to 5.8% in 1998. Multiresistance, defined as resistance to norfloxacin and at least two of the other three antibiotics, increased from 0.5% in 1989 to 4. 0% in 1998. Multivariate analysis of the norfloxacin resistance demonstrated that this yearly increase (the odds ratio was 1.0 in 1989, 1.6 in 1992, 2.9 in 1995 and 6.1 in 1998) was independent of other determinants of resistance to norfloxacin, such as age, gender and origin of the isolate. Analysis of strata, classified by year, age and gender, demonstrated an association between prescription of fluoroquinolones (defined daily doses per case of UTI) and resistance to norfloxacin in E. coli (P < 0.001). There was no significant association with the prescription of nitrofuran derivatives (nitrofurantoin) and trimethoprim with or without sulphamethoxazole. The yearly increase of resistance to fluoroquinolones in E. coli from UTI may stem from increased prescription of fluoroquinolones for UTI. Resistance of E. coli to these agents is likely to increase further as fluoroquinolone use increases in future.

Three cases of serious infection caused by Aerococcus urinae.

Three cases of serious infection caused by Aerococcus urinae are presented: a patient with endocarditis and two patients with soft-tissue infection (phlegmon and balanitis respectively). The literature on Aerococcus urinae infections is reviewed and the antibiotic therapy discussed. Aerococcus urinae is a pathogen isolated primarily from urine specimens of elderly patients with local or systemic predisposing conditions. Most infections are mild, but serious infections such as endocarditis and septicemia/urosepsis have been described. Penicillin or ampicillin in combination with an aminoglycoside and close monitoring of the patient's clinical status and laboratory results would seem to be the best strategy for management of cases of serious infection.

Clinical spectrum of infections due to the newly described Actinomyces species A. turicensis, A. radingae, and A. europaeus.

Over a 7-year period, we isolated 294 Actinomyces-like organisms (ALOs) which were not clearly identifiable. Using well-defined probes coding for sequences specific for recently described Actinomyces species (A. turicensis, A. radingae, and A. europaeus), we were able to identify 128 strains. The majority belonged to the A. turicensis species. A. radingae was found only in patients with skin-related pathologies. A. europaeus was also detected in patients with urinary tract infections. The main sources of A. turicensis were genital infections, followed by skin-related and urinary tract infections. Additional clinical pictures were appendicitis, cholecystitis, ear, nose, and throat infections, and bacteremia. In a small number of patients these ALOs were found as the only pathogen. Strains of the three species were tested by two widely used biochemical identification methods. A. turicensis was easily identifiable by both these methods. We conclude that these ALOs are not infrequent pathogens and are found in a wide range of human infections. At least A. turicensis is easily identifiable by clinical diagnostic laboratories.

Occurrence of extended-spectrum betalactamases (ESBL) in Dutch hospitals.

The prevalence of ESBL was determined among isolates of Escherichia coli (n = 571) and Klebsiella spp. (n = 196) collected during a 1-week study period in 8 university and 3 large regional laboratories all over the Netherlands. 18 isolates were positive for at least one of the screening tests used, i.e., VITEK-ESBL, E-test ESBL and MIC ratio of ceftazidime/ceftazidime-clavulanic acid, cefotaxime/cefotaxime-clavulanic acid. In 5 of these 18 putative ESBLs no betalactamase production was detectable. A TEM type was found in three E. coli and two Klebsiella spp. An SHV type was present in five Klebsiella spp. In one E. coli and one Klebsiella pneumoniae both enzymes were present. In one Klebsiella oxytoca neither of the two enzymes was present. Using PCR for both ESBL TEM and ESBL SHV, an SHV ESBL was found in one E. coli and four Klebsiella isolates. The mutations at position 238 and 240 were already described. In one E. coli isolate a TEM ESBL was found with three mutations, at position 21, 164 and 265. These mutations were already described in other ESBLs but not in this combination suggesting a new TEM ESBL. The overall prevalence of ESBL producing E. coli and Klebsiella spp. was less than 1% (6 out of 767).

Characterization of Actinomyces turicensis and Actinomyces radingae strains from human clinical samples.

Whole-organism protein electrophoresis was used to compare and group unidentified coryneform bacteria resembling Gardnerella vaginalis and various Actinomyces and Arcanobacterium species. The obtained clusters of strains were further characterized by whole-cell fatty acid analysis and a variety of biochemical tests. Species-specific oligonucleotide probes based on 16S rRNA gene sequences were designed. The results demonstrate that the majority of the isolates belonged to Actinomyces turicensis; the other strains belonged to Actinomyces radingae. The descriptions of both species are emended.

Resistance of staphylococci in The Netherlands: surveillance by an electronic network during 1989-1995.

An electronic surveillance network for monitoring antibiotic resistance in The Netherlands has been in operation since 1989. Seven public health laboratories participate and the system covers about 25% of all bacteriological determinations in The Netherlands. This paper reports the results of staphylococci isolated in the period 1989-1995. About 0.3% of the Staphylococcus aureus isolates in the study period were resistant to methicillin. This low percentage may be due to the restrictive use of antibiotics and to strict isolation measures aimed at eradicating methicillin-resistant S. aureus. Low frequencies of resistance among methicillin-resistant S. aureus were found for vancomycin (0%), chloramphenicol (11%), cotrimoxazole (11%), mupirocin (3% low-level resistance) and fusidic acid (7%). Twenty-one percent of the coagulase-negative staphylococci were resistant to methicillin. Low frequencies of resistance among these methicillin-resistant coagulase-negative staphylococci were those to vancomycin (0.4%), nitrofurantoin (2%), doxycycline (20%) and amikacin (20%). Coagulase-negative staphylococci from cerebrospinal fluid, blood and skin were less often resistant to quinolones than isolates from respiratory tract, faeces and urine. A significant increase in resistance of coagulase-negative staphylococci to methicillin, erythromycin, gentamicin and ciprofloxacin was observed in the investigated period but the resistance to doxycycline and co-trimoxazole decreased in the last few years. To confirm the determination of methicillin resistance and coagulase production, a PCR method was developed which detects both the mecA and the coagulase gene. The results of the PCR method correlated well with the methicillin MIC as determined by an agar-dilution method.

Pitfalls and fallacies of cat scratch disease serology: evaluation of Bartonella henselae-based indirect fluorescence assay and enzyme-linked immunoassay.

The diagnostic value of the detection of immunoglobulin G (IgG) and IgM by Bartonella henselae-based indirect fluorescence assay (IFA) and enzyme-linked immunoassay (EIA) for the diagnosis of cat scratch disease (CSD) was evaluated. The IFA was performed either with B. henselae that was cocultivated for a few hours with Vero cells or with noncocultivated B. henselae as the antigen. Additionally, the performance of a Bartonella PCR hybridization assay based on the 16S rRNA gene was determined and compared with those of the serologic assays. The study group consisted of 45 patients suspected of suffering from CSD by fulfilling one or more of the classical criteria. The specificities of the immunoassays were set at > or = 95% by analysis of sera from 60 healthy blood donors. It is shown that the sensitivities of the IgG assays are very low (40.9% for the IFA with noncocultivated B. henselae as antigen) and that those of the IgM assays are higher (71.4% for the EIA) for patients who fulfilled two or more criteria for CSD. The IgM EIA showed the highest sensitivity: 71.4% in patients with two or more criteria for CSD and 80.6% for patients with a positive Bartonella PCR result. The results indicate that the specificities of both IFA and EIA IgG serologies and the sensitivity of the IFA IgM serology need to be improved. The PCR hybridization assay showed a sensitivity of 86.4% for patients who fulfilled two or more criteria for CSD and 100% for seven patients who fulfilled three or more criteria. The kinetics of IgG and IgM antibody production were studied in 18 patients with CSD on the basis of a positive B. henselae IFA IgM serology. The results indicate that there is no standard course of anti-B. henselae IgG and IgM production in patients with CSD, because some patients produced high levels of both IgG and IgM, others produced only high levels of IgM, and a few patients produced only low levels of antibodies.

Actinomyces europaeus sp. nov., isolated from human clinical specimens.

Ten strains of a hitherto undescribed catalase-negative, facultatively anaerobic, coryneform bacterium were isolated or collected by workers at three European clinical bacteriology laboratories or reference centers. These strains were isolated from humans, and most came from abscess material. Biochemical and chemotaxonomic characterization revealed that the strains belonged to the genus Actinomyces. The phenotypic features of the 10 strains were incompatible with the descriptions of the previously established Actinomyces species. A comparative 16S rRNA gene sequence analysis demonstrated that the previously undescribed strains constitute a new line in the genus Actinomyces. The name Actinomyces europaeus sp. nov. is proposed for these clinical isolates. The type strain is CCUG 32789A.

Urinary tract infections with Aerococcus urinae in the south of The Netherlands.

Aerococcus urinae is an uncommon urinary tract pathogen that causes infections predominantly in elderly persons with local or general predisposing conditions. During a one-year study, the clinical features of Aerococcus urinae urinary tract infections (> or = 10(5) cfu/ml) were investigated in two large medical microbiology laboratories in the Netherlands. The incidence of Aerococcus urinae urinary tract infections ranged between 0.31 and 0.44% for the two laboratories. The median age (range 35-95 years) of patients with this infection was 82.5 years for women and 77.5 for men. Men had significantly (p < 0.01) more local predisposing conditions than did women. Underlying systemic diseases such as diabetes mellitus, malignancy, and dementia were found in 67.5% of patients. Most patients (97.5%) had the classic signs of a urinary tract infection, but none of them developed serious symptoms. All isolates tested were susceptible to penicillin, amoxicillin, and nitrofurantoin, 78.3% were susceptible to norfloxacin, and all were resistant to sulfonamides. The majority of patients were treated with amoxicillin, amoxicillin with clavulanic acid, or norfloxacin.

[Intravenously-administered immunoglobulins as first-choice agent in juvenile dermatomyositis]. / Intraveneus toegediend immunoglobuline als middel van eerste keuze bij juveniele dermatomyositis.

Dermatomyositis is an acquired disease characterised by symmetric predominantly proximal muscle weakness of the arms and legs, and misery. It may be associated with myalgia and there is often a characteristic rash. The mainstay of therapy is corticosteroids. Recently efficacy of intravenous immunoglobulin (IVIg) in chronic refractory dermatomyositis was reported. Because corticosteroids can cause serious side effects, we treated a seven-year-old girl suffering from dermatomyositis with IVIg as initial therapy. After two courses of IVIg infusions at a dose of 0.4 g/kg/day for five consecutive days, the patient made a rapid and complete recovery. This case shows that IVIg may be effective as initial therapy in patients with dermatomyositis. Whether IVIg is really a better treatment than corticosteroids should be investigated in a randomised study.

Hepatitis C virus infection in African patients with liver cirrhosis or primary hepatocellular carcinoma.

We have studied the prevalence of hepatitis C virus (HCV) infection in Rwandan patients with histologically proven liver cirrhosis (LC) or primary hepatocellular carcinoma (HCC). Anti-HCV antibodies were determined by using a second-generation test, with a line immunoassay for structural and non-structural antigens as confirmation. Seventy-nine patients with LC, 26 with HCC, and 54 voluntary blood donors as controls were evaluated. Anti-HCV antibodies were more prevalent in LC patients (48%) and in HCC patients (38%) than in the controls (17%; difference, p = 0.0001 and p = 0.03, respectively). Eighty-four per cent of LC patients and 54% of HCC patients were HBsAg-negative. The prevalence of anti-HCV antibodies was significantly higher for LC and HCC patients who had been in contact with HBV but who had no persistent HBV infection (p < 0.05). We conclude that HCV infection is common in Rwanda and is linked to LC and HCC.

Development of an antibody-capture IgM-enzyme-linked immunosorbent assay for diagnosis of acute Epstein-Barr virus infections.

An anti-EBV IgM-ELISA was developed using the antibody-capture principle, to be used for the diagnosis of acute infectious mononucleosis (IM). The test was based on anti-human IgM-coated microtiter plates; nuclei of EBV producer cells were used for antigen; conjugate was prepared by labeling sheep anti-EBV IgG with horseradish peroxidase. The specificity of the anti-EBV IgM-ELISA was studied with a panel of sera from acute infections with hepatitis A virus, rubella virus, Toxoplasma gondii and cytomegalovirus, and sera positive for rheumatoid factors, positive for antinuclear antibodies, as well as with sera from normal blood donors and pregnant women. Specificity in these panels was 98.4%. In a clinical study with 449 sera from patients with IM-like symptoms, 109 of 109 confirmed patients were detected by the anti-EBV IgM-ELISA. Specificity of the anti-EBV IgM-ELISA in this clinical study was 99.7%. The anti-EBV IgM-ELISA detected several acute EBV patients who had negative heterophile antibody titers.

Ability of enzyme-linked immunosorbent assays to detect early immunoglobulin G antibodies to Toxoplasma gondii.

Two ELISA procedures, one using sonicated antigen coated with carbonate buffer and the other formalin fixed trophozoites with dry coating, differ in their ability to detect early antibodies in toxoplasmosis. In order to identify factors responsible for this difference, seven ELISA systems differing from each other in antigen used and/or coating procedure were compared. Both fixation of the trophozoites with formalin and air-drying of the antigen in the microtiterplate were important factors determining the ability of the assay to detect IgG antibodies in the early stage of infection. Differences in the results of the two ELISA procedures can be used to distinguish between the acute and chronic stages of infection.