Pertussis disease burden in the household: how to protect young infants.

BACKGROUND: We conducted a population-based, nation-wide, prospective study to identify who introduced pertussis into the household of infants aged 6 months admitted to the hospital for pertussis in the Netherlands. METHODS: During the period 2006-2008, a total of 560 household contacts of 164 hospitalized infants were tested by polymerase chain reaction, culture, and serological examination to establish Bordetella pertussis infection. Clinical symptoms and vaccination history were obtained by a questionnaire submitted during sample collection and 4-6 weeks afterwards. RESULTS: Overall, 299 household contacts (53%) had laboratory-confired pertussis; 159 (53%) had symptoms compatible with typical pertussis infection, and 42 (14%) had no symptoms. Among children vaccinated with a whole-cell vaccine, 17 (46%) of 37 had typical pertussis 1-3 years after completion of the primary series, compared with 9 (29%) of 31 children who had been completely vaccinated with an acellular vaccine. For 96 households (60%), the most likely source of infection of the infant was established, being a sibling (41%), mother (38%), or father (17%). CONCLUSIONS: If immunity to pertussis in parents is maintained or boosted, 35%-55% of infant cases could be prevented. Furthermore, we found that, 1-3 years after vaccination with whole-cell or acellular vaccine, a significant percentage of children are again susceptible for typical pertussis. In the long term, pertussis vaccines and vaccination strategies should be improved to provide longer protection and prevent transmission.

Evaluation of Legionella V-TesT for the detection of Legionella pneumophila antigen in urine samples.

We evaluated a new immunochromatographic assay (Legionella V-TesT, Coris BioConcept, Gembloux, Belgium) for its ability to detect Legionella pneumophila serogroup 1 antigen in urine. Test devices were read at various time points to determine the optimum incubation time regarding performance. The results were compared with those obtained with the BinaxNOW urinary antigen test. The sensitivity and specificity were 82.2% and 98.6%, respectively, for the Legionella V-TesT and 83.9% and 100%, respectively, for the BinaxNOW urinary antigen test after 15 min of incubation. When tests were examined after 60 min, the sensitivity for both tests increased to 91.5%.

Evaluation of the Oxoid Xpect Legionella test kit for detection of Legionella pneumophila serogroup 1 antigen in urine.

We evaluated a new immunochromatographic assay (Oxoid Xpect Legionella test kit) for the ability to detect Legionella pneumophila serogroup 1 antigen in urine. The results were compared with those obtained with the Binax NOW urinary antigen test by following the manufacturers' instructions. The sensitivities and specificities were estimated to be 89 and 100%, respectively, for the Oxoid Xpect Legionella test kit and 86 and 100%, respectively, for the Binax NOW test.

Pelvic abscess caused by Arcanobacterium haemolyticum mimicking a soft tissue tumour.

Arcanobacterium haemolyticum has usually been isolated in cases of pharyngitis and wound infections. Rarely it has been reported to cause deep tissue infections. Here, a case of a 71-year-old-male, who developed a pelvic abscess due to A. haemolyticum that initially was thought to be a malignant tumour, is described.

Evaluation of the SAS Legionella Test, a new immunochromatographic assay for the detection of Legionella pneumophila serogroup 1 antigen in urine.

This study evaluated a new immunochromatographic assay (SAS Legionella Test) for its ability to detect Legionella pneumophila serogroup 1 antigen in urine. Results were compared with those obtained using the Binax Now urinary antigen test. Sensitivity and specificity were estimated as 82.9% and 99.0%, respectively, for the SAS Legionella Test, and 91.4% and 100%, respectively, for the Binax Now urinary antigen test. The sensitivity of both tests increased to 97.1% (p 0.009) and 94.2% (p 0.7), respectively, if the tests were examined after 1 h.

Serological testing for Bartonella henselae infections in The Netherlands: clinical evaluation of immunofluorescence assay and ELISA.

Cat-scratch disease (CSD), caused by Bartonella henselae infection, can mimic malignancy and can manifest atypically. Reliable serological testing is therefore of great clinical importance. The diagnostic performance of immunofluorescence assay (IFA) and ELISA was evaluated in a group of Dutch patients with proven CSD (clinical diagnosis confirmed by PCR). Sera of 51 CSD patients and 56 controls (patients with similar symptoms, but who were B. henselae PCR-negative and had an alternative confirmed diagnosis) were tested for anti-B. henselae IgM and IgG by IFA and ELISA. A commercially available IFA test for IgM had a sensitivity of 6%. In-house assays for IgM showed specificities of 93% (IFA) and 91% (ELISA), but with low sensitivities (53% and 65%, respectively). With a specificity of 82% (IFA) and 91% (ELISA), in-house IgG testing showed a significantly higher sensitivity in IFA (67%) than in ELISA (28%, p <0.01). Sensitivity was higher for genotype I (38-75%) than for genotype II (7-67%) infections, but this was only statistically significant for IgG ELISA (p <0.05). In conclusion, detection of IgM against B. henselae by in-house ELISA and IFA was highly specific for the diagnosis of CSD. The high seroprevalence in healthy individuals limits the clinical value of IgG detection for diagnosing CSD. Given the low sensitivity of the serological assays, negative serology does not rule out CSD and warrants further investigation, including PCR. Adding locally isolated (e.g., genotype II) B. henselae strains to future tests might improve the sensitivity.

No evidence of Legionella infection in general practice patients presenting with acute respiratory infections in The Netherlands.

The role of Legionella spp. in the aetiology of acute respiratory infections (ARIs) is largely unknown. In this case-control study, conducted in a general practitioner setting during 2000 and 2001, nose and throat samples from patients presenting with ARIs (n = 230) and controls (n = 200) were analysed for the presence of Legionella spp. by real-time PCR. Legionella DNA was not detected in any of the cases or controls. Thus, Legionella spp. do not seem to play a role in patients presenting with ARIs, nor were they present in patients who visited their general practitioner for complaints other than ARIs.

Community-acquired pathogens associated with prolonged coughing in children: a prospective cohort study.

A 2-year prospective study was performed of children with prolonged coughing to investigate the frequency of different respiratory pathogens, the rate of mixed infections, and possible differences in severity of disease between single and mixed infections. Sera from 135 children (136 episodes of prolonged coughing lasting 1-6 weeks) were tested for antibodies to different viruses and bacteria. Swabs were taken for culture and PCR to detect different viral and bacterial pathogens. One or more pathogens were found in 91 (67%) patients. One infectious agent was found in 49 (36%) patients, two agents in 35 (26%) patients, and more than two agents in seven (5%) patients. The most frequent pathogens encountered were rhinovirus (n = 43; 32%), Bordetella pertussis (n = 23; 17%) and respiratory syncytial virus (n = 15; 11%). The most frequent mixed infection was B. pertussis and rhinovirus (n = 14; 10%). No significant differences in clinical symptoms were observed between patients with or without pathogens; however, patients with mixed infections were significantly older. There was a strong seasonal influence on the number of infections, but not on the number of mixed infections. In children with prolonged coughing, there was a high frequency of mixed infections regardless of the season. However, mixed infection was not associated with increased disease severity. No clinical symptoms were found that allowed discrimination between specific pathogens.

Erythromycin resistance in the commensal throat flora of patients visiting the general practitioner: a reservoir for resistance genes for potential pathogenic bacteria.

The prevalence and mechanism of erythromycin resistance in commensal throat streptococci was determined from October 2000 until December 2002 as part of an ongoing study of the NIVEL in general practice patients (N=678). Resistance prevalence for 1mg/L and 16 mg/L erythromycin was 57% and 20%, respectively. The percentage of total commensal flora resistant within each patient ranged from 1% to 100% (median, 1%). mefA was predominantly found among isolates on the 1mg/L plates, and ermB was found in 64% of the isolates on the 16 mg/L plates. Erythromycin resistance was transferred from a commensal isolate to Streptococcus pneumoniae with a frequency of 1 x 10(-9). Commensal streptococci of general practice patients in The Netherlands form a large reservoir of transferable erythromycin resistance (genes) for potential pathogenic microorganisms.

Chlamydia pneumoniae, systemic inflammation and the risk of venous thrombosis.

Inflammatory mediators are involved in activation of the coagulation system, and elevated plasma concentrations of IL-6 and IL-8 are associated with an increased risk of venous thrombosis. Using serologic and molecular biologic tests, we investigated in a case-control study on patients with recurrent venous thrombosis the association between Chlamydia (C) pneumoniae and venous thrombosis and we evaluated the relation between C. pneumoniae serology and the cytokines IL-6 and IL-8. The presence of C. pneumoniae antibody titers > or = 1:16 was not associated with an increased risk of venous thrombosis (odds ratio 0.8 95% CI, 0.4-1.7). Circulating C. pneumoniae-DNA was detected in only one patient and two control subjects. IgG antibody titers against C. pneumoniae were not correlated with the concentrations of IL-6 and IL-8. These results indicate that the inflammatory process shown in patients with venous thrombosis is not related to C. pneumoniae.

Extraction of Chlamydia pneumoniae DNA from vascular tissue for use in PCR: an evaluation of four procedures.

The objective of this study was to compare four procedures for Chlamydia pneumoniae DNA extraction from vascular tissue. The NucliSens Kit, the QIAamp tissue DNA MiniKit, buffer-saturated phenol and the Geneclean II Kit were evaluated, based on the yield of recovered DNA, using PCR to detect C. pneumoniae in vascular tissue. The QIAamp tissue procedure had the highest detection level (0.004 inclusion-forming units/sample). All methods, except NucliSens (70 min), had a short handling time (30-40 min). Costs varied from 0.5 to 3.2 Euro.

Nosocomial influenza infection among post-influenza-vaccinated patients with severe pulmonary diseases.

OBJECTIVES: To report an outbreak of nosocomial influenza in thirteen out of twenty-two admitted patients suffering from severe lung emphysema. METHODS: Acute-phase and convalescent serum samples of nine patients were collected. An antihaemagglutinin assay was performed to detect a rise in antibodies against influenza A virus. Further information about vaccination history of the patients and healthcare workers was included. RESULTS: The majority of these twenty-two patients was vaccinated with a trivalent vaccine six months earlier. The immunological response showed that the influenza A (H3N2) strain which caused these infections is similar to the vaccine strain A/Sydney/5/97. CONCLUSIONS: The staff of our institute which was not systematically vaccinated may have been the source of infection. The time elapsed between the vaccination and the infection is the probable explanation of this event.

Tubal factor pathology caused by Chlamydia trachomatis: the role of serology.

One of the causes of infertility in females is tubal pathology as a result of pelvic inflammatory disease (PID) caused by Chlamydia trachomatis (Ct). Diagnosis and identification of patients is hampered by the lack of rapid, easy, sensitive and specific methods. The introduction of Ct-specific enzyme-linked immunosorbent assay (ELISA) tests, based upon synthetic peptides may subsequently increase the sensitivity and specificity for the detection of tubal factor infertility caused by Ct. In order to determine the value of these tests for serological diagnosis of Ct infections, we evaluated several commercially available assays (C. trachomatis enzyme immunoassay (EIA), Labsystems (CtL); SeroCT, Savyon (CtS); pELISA, Medac (CtMp); and a reference assay rELISA, Medac (CtMr)) in two study populations. The first group consisted of 134 female patients with infertility problems. Tubal factor infertility was observed in 85 of these patients (63%). A higher % positivity was found for Ct-specific IgG for the CtL, CtS and CtMp, 41% vs 10%, 57% vs 18% and 55% vs 25% respectively as compared to patients with infertility due to other problems. A similar trend was observed for Ct-specific IgA. The specificity of Ct-specific IgA and IgG in this patient group varied between 92 to 98% and 76 to 90%, respectively. The second group consisted of 107 consecutive gynaecology patients with fertility problems or suspected PID. In this particular patient group, the specificity of the peptide based tests were around 80% and 90% for Ct-specific IgA and 75% and 85% for Ct-specific IgG, respectively. The negative predictive values exceeded 90%, while the positive predictive values varied from 30% to 47% for Ct-specific IgA and was around 30% for Ct-specific IgG. Testing Ct-specific IgG had no additional value above Ct-specific IgA alone. We conclude that the new synthetic peptide-based EIA tests are able to detect species-specific antibodies, which are correlated to (active) infection, and that in particular IgA may be useful in the serodiagnosis of tubal factor infertility caused by C. trachomatis, and will contribute in simplifying the work-up in patients with infertility.

Sensitivity and specificity of three new commercially available Chlamydia trachomatis tests.

In order to determine the value of new Chlamydia trachomatis (Ct) specific tests for routine serological diagnosis of Ct infections, we evaluated several commercially available assays (C. trachomatis enzyme immunoassay (EIA), Labsystems (CtL); SeroCT, Savyon (CtS); pELISA, Medac (CtMp)) in various study populations. The prevalence of C. trachomatis-specific IgA antibodies in a blood donor population (n = 443) as determined by the peptide based tests CtL, the CtS and the CtMp was 5%, while for IgG antibodies this was 6% (CtL and CtS) and 12% (CtMp) respectively. Prevalence was negatively correlated with age, concording with C. trachomatis specificity. None of the three tests showed significant titre rises in serum samples taken from patients with a proven infection of Chlamydia pneumoniae (n = 22), indicating species-specificity for all three tests. In patients with a polymerase chain reaction proven (n = 324) Ct infection, 75%, 70% and 68% were positive for IgG and 45%, 38% and 47%, positive for IgA as determined by the CtMp, CtL and CtS respectively. We conclude that the new synthetic peptide-based EIA tests are able to detect species-specific Ct antibodies, which are strongly correlated to (active) infection.

Rapid diagnosis of herpes encephalitis by enzyme immuno-assay.

Five cases of encephalitis caused by herpes simplex virus (HSV) are described. HSV-specific IgM and IgM antibodies were detected in cerebrospinal fluid and serum by use of antibody-capture noncompetitive enzyme-linked immunosorbent assay. The tests seem suitable for rapid diagnosis in the second week after onset of neurological symptoms. The clinical importance of early diagnosis and therapy in patients of HSV encephalitis has been discussed.

[Is Chlamydia TWAR of significance in The Netherlands?]. / Heeft Chlamydia TWAR betekenis in Nederland?

Chlamydia TWAR is as a newly recognised organism that causes respiratory tract infection with human-to-human transmission. Our sero-epidemiological study showed that in The Netherlands prevalence of TWAR antibodies is very low in children under the age of five years, increases after beginning of school age until adolescence, and remains high (80%) during adulthood. It is known that Chlamydia TWAR can cause acute lower respiratory tract infection. However, the exact clinical spectrum within the various groups of patients still needs to be defined.

[Maxillary sinusitis in children]. / Sinusitis maxillaris bij kinderen.

In children with nasal discharge the distinction between the group with a diagnosis of rhinitis and the group with a diagnosis of sinusitis was vague. Anamnestic data and findings at physical examination were not significantly different. There was hardly any relation between radiographic and echographic findings, the nature of the irrigation fluid, and isolation of pathogenic or non-pathogenic bacteria from cultures of nasal or sinus secretions. In our opinion there is no clearcut difference between rhinitis and simple sinusitis (i.e. mucositis of the maxillary sinus without empyema). Nor is this therapeutically relevant because both conditions are mostly features of a respiratory tract infection. It is therapeutically important, however, to differentiate between sinusitis without and sinusitis with empyema. The latter requires special treatment. None of the patients studied had a sinusitis with empyema. When a child has running nose there is no reason to look for a sinusitis when there are no clinical symptoms of empyema.

[Aeromonas species as cause of diarrhea and infections outside the gastrointestinal tract in The Netherlands]. / Aeromonas-species als verwekker van diarree en infecties buiten het maag-darmkanaal, in Nederland.

In a national survey in the period May 1986-December 1987, Aeromonas was isolated from 277 out of 16,857 (1.6%) samples of watery, bloody or mucous stool, from patients with diarrhoea. There was a clear seasonal pattern (less than 1% in winter, up to 3% in summer). A. caviae was isolated most frequently (49%), followed by A. sobria (35%) and A. hydrophila (15%). Some non-identifiable strains were isolated as well. Aeromonas were isolated in particular from faeces of patients aged over 70 years (predominantly A. sobria) or under 5 years (predominantly A. caviae). In 67% Aeromonas was isolated as the only possible bacterial cause of diarrhoea, but in 33% other enteropathogenic bacteria were found as well (17% Campylobacter, 14% Salmonella, 2% Shigella). In addition, all Aeromonas isolates were collected which were obtained in normal diagnostic activities in the participating laboratories, among others from blood, from pus or wound fluid, from faecal samples which did not meet the above mentioned criteria. A. caviae was the dominant species in 'other' faeces and 'various' body sites but was not isolated from blood. The results of this study do not indicate that routine examinations for Aeromonas in faeces of patients with diarrhoea are necessary.

Influenza A/H1N1 vaccination response is inadequate in down syndrome children when the latest cut-off values are used.

We determined the response of 48 Down syndrome children to 2 doses of influenza A/H1N1 vaccination. Ninety-two percent of the children reached the previously defined protective level (hemagglutination-inhibition titer ≥1:40), but only 27% of the children reached the level of ≥1:110 which was recently described to predict the conventional 50% clinical protection rate in children. Further studies, and potentially adaptations of the schedule, are needed.

Detection of enterovirus RNA in cerebrospinal fluid: comparison of two molecular assays.

Enterovirus (EV) and human parechovirus (HPeV) are a major cause of infection in childhood. A rapid diagnostic test may improve the management of patients with EV and HPeV infection. The aim of this study is to evaluate the performance of the GeneXpert enterovirus assay (GXEA) for detection of EV RNA compared to a user-developed reverse-transcriptase (RT) quantitative real-time PCR (qPCR) in routine clinical practice. Also a RT-qPCR assay for detection of HPeV RNA in different clinical samples was developed and evaluated. Cerebrospinal fluid (CSF) from 232 patients suspected for meningitis was collected and tested for EV and HPeV using RT-qPCR assays. In parallel an aliquot of the samples was tested using the GXEA and viral culture. EV RNA was detected in 22 (19.0%) and 28 (24.1%) of 116 samples using the GXEA and RT-qPCR assay, respectively. EV was isolated from 10 of 116 (8.6%) samples by viral culture. GXEA had a sensitivity, specificity, positive predictive value and negative predictive value of 82.1%, 100%, 100% and 96.2%, respectively. In this study, molecular assays were superior to viral culture for detecting EV RNA in CSF. GXEA showed a high specificity but a lower sensitivity for the detection of EV RNA compared to the RT-qPCR assay.