The need for European professional standards and the challenges facing clinical microbiologyHumphreys, H.; Nagy, E.; Kahlmeter, G.; Ruijs, G. J. H. M.
doi: 10.1007/s10096-010-0906-2pmid: 20333424
Microorganisms spread across national boundaries and the professional activities of clinical (medical) microbiologists are critical in minimising their impact. Clinical microbiologists participate in many activities, e.g. diagnosis, antibiotic therapy, and there is a need for a set of professional standards for Europe with a common curriculum, to build upon the current strengths of the specialty and to facilitate the free movement of specialists within the European Union. Such standards will also better highlight the important contribution of clinical microbiologists to healthcare. There is a move to larger centralised microbiology laboratories often located off-site from an acute hospital, driven by the concentration of resources, amalgamation of services, outsourcing of diagnostics, automation, an explosion in the range of staff competencies and accreditation. Large off-site centralised microbiology laboratories are often distant to the patient and may not facilitate the early detection of microbial spread. Ultimately, the needs of patients and the public are paramount in deciding on the future direction of clinical microbiology. Potential conflicts between integration on an acute hospital site and centralisation can be resolved by a common set of professional standards and a team-based approach that puts patients first.
Skin bacteria after chlorhexidine exposure—is there a difference in response to human β-Defensin-3?Reichel, M.; Heisig, A.; Heisig, P.; Kampf, G.
doi: 10.1007/s10096-010-0904-4pmid: 20339890
We investigated whether exposure to sub-lethal concentrations of chlorhexidine digluconate (CHG) changed the response of five Staphylococcus spp. to human β-Defensin-3 (hBD-3). The change in response for each strain was determined in vitro with time–kill experiments in suspension by comparing the mean log10 reduction caused by hBD-3 at 1.5 and 3 h in exposed and non-exposed bacteria. The identity of staphylococcal species was verified by DNA sequence homology in the gyrA genes in comparison with reference strains. Baseline sub-lethal concentrations allowing visible bacterial growth were between 0.0625 and 0.25 μg/ml. Sub-lethal CHG concentrations increased within 3 days in two isolates. For S. capitis 19/2, CHG-exposed cells were less susceptible to 0.5 μg/ml hBD-3 (log10 reduction 0.78 versus 2.06 at 1.5 h; p < 0.001; t-test). For S. aureus, however, CHG-exposed cells were more susceptible to 1 μg/ml hBD-3. The observed changes between CHG-exposed and non-exposed cells did not indicate a general trend in response to hBD-3. Overall, we found no consistent evidence that 3 days of exposure to CHG changed the response of five Staphylococcus spp. to hBD-3. The use of CHG for skin antisepsis is, based on our data, unlikely to change the natural defence activity of hBD-3.
Eradication of the commensal intestinal microflora by oral antimicrobials interferes with the host response to lipopolysaccharideUmenai, T.; Hirai, H.; Shime, N.; Nakaya, T.; Asahara, T.; Nomoto, K.; Kita, M.; Tanaka, Y.; Imanishi, J.
doi: 10.1007/s10096-010-0905-3pmid: 20300949
The host components and commensal microorganisms of the intestinal microenvironment play roles in the development and maintenance of the host defence. Recent observations have suggested that toll-like receptors (TLRs) are involved in the recognition of innate immunity against intestinal microbes. However, little is known regarding the role of TLR in the maintenance of systemic host defence by intestinal microorganisms. We studied the expression and function of TLR4 and TLR2 on alveolar and peritoneal macrophages in mice after 3 weeks of oral administration of streptomycin and cefotaxime. After active treatment, the intestinal microorganisms were nearly completely eradicated, and the surface expression of TLR4 and TLR2 on the peritoneal macrophages was prominently downregulated. When the actively treated mice were challenged with lipopolysaccharide (LPS), a TLR4 ligand, the host response was markedly impaired. Our results suggest that the oral administration of antimicrobials downregulates the expression of surface TLR on the peritoneal macrophages and modulates the host immune responses against LPS by modifying the intestinal environment.
T regulatory cells and Th1/Th2 cytokines in peripheral blood from tuberculosis patientsHe, X.-Y.; Xiao, L.; Chen, H.-B.; Hao, J.; Li, J.; Wang, Y.-J.; He, K.; Gao, Y.; Shi, B.-Y.
doi: 10.1007/s10096-010-0908-0pmid: 20306324
About 10% of people infected with Mycobacterium tuberculosis develop active tuberculosis (TB), and Th1 effector cells and Th1 cytokines play key roles in controlling M. tuberculosis infection. Here, we hypothesise that this susceptibility to M. tuberculosis infection is linked to increased T regulatory (Treg) cells and Th2 cytokines in TB patients. To test this, we recruited 101 participants (71 TB patients, 12 non-TB pulmonary diseases and 18 healthy subjects) and investigated Treg cells and Th1/Th2 cytokines in peripheral blood. CD4+CD25+ T cells and CD4+CD25+FoxP3+ T cells significantly increased and IL-5 dramatically decreased in TB patients relative to healthy subjects. CD8+CD28− T cells, IFN-γ, TNF-α, IL-10 and IL-4 significantly increased in patients with culture and sputum smear-positive pulmonary TB (PTB(+)) compared with healthy subjects. CD4+CD25+FoxP3+ and CD8+CD28− T cells significantly decreased in PTB(+) after one month of chemotherapy. CD4+, CD4+CD25+ and CD8+CD28+ T cells significantly increased in extra-pulmonary TB patients after one month of chemotherapy. These findings suggest that M. tuberculosis infection induces circulating CD4+CD25+FoxP3+ and CD8+CD28− T cell expansion, which may be related to the progression of M. tuberculosis infection, and that the balance between effector immune responses and suppression immune responses is essential to control M. tuberculosis infection.
Characterisation of invasive Haemophilus influenzae isolates in Slovenia, 1993–2008Kastrin, T.; Paragi, M.; Kolman, J.; Čižman, M.; Kraigher, A.; Gubina, M.
doi: 10.1007/s10096-010-0910-6pmid: 20306323
The objectives of our study were to describe the epidemiology of invasive Haemophilus influenzae disease from 1993 to 2008 in Slovenia, a country with routine H. influenzae serotype b (Hib) conjugate vaccination since the year 2000. A total of 292 isolates of H. influenzae, recovered from a normally sterile site, were collected in the study period. The isolates were serotyped by slide agglutination and antibiotic susceptibility was determined. One hundred and eight isolates received after the year 2000 were serotyped by slide agglutination and by polymerase chain reaction (PCR) capsule typing, and both methods were compared. After the introduction of the routine Hib vaccination, the incidence of H. influenzae disease in children under the age of 5 years has decreased by 87.6% and type b was replaced by non-typeable H. influenzae as the predominant serotype. The proportion of serotype b decreased from 85.3% in the pre-vaccination period to 13.0% in the vaccination period and the proportion of non-capsulated isolates increased from 12.0 to 85.2%. The study of genetic relatedness by pulsed-field gel electrophoresis (PFGE) demonstrated that the isolates of serotypes b and f were genetically homogeneous within the serotype. The results of our national surveillance showed that the vaccine has been very efficient in preventing Hib invasive disease in Slovenia. Nevertheless, we see a need for further monitoring of invasive H. influenzae infections at a national level.
Comparison of five commercial serological tests for the detection of anti-Chlamydia trachomatis antibodiesBaud, D.; Regan, L.; Greub, G.
doi: 10.1007/s10096-010-0912-4pmid: 20349260
Screening for Chlamydia trachomatis-specific antibodies is valuable in investigating recurrent miscarriage, tubal infertility and extrauterine pregnancy. We compared here the performance of immunofluorescence (IF) to four other commercial tests in detecting IgG antibodies directed against C. trachomatis: two enzyme-linked immunosorbent assays (ELISAs) using the major outer membrane protein (MOMP) as the antigen, commercialised respectively by Medac and R-Biopharm (RB), one ELISA using the chlamydial heat shock protein 60 (cHSP60) as the antigen (Medac), as well as a new automated epifluorescence immunoassay (InoDiag). A total of 405 patients with (n = 251) and without (n = 154) miscarriages were tested by all five tests. The prevalence of C. trachomatis-specific IgG antibodies as determined by the IF, cHSP60-Medac, MOMP-Medac, MOMP-RB and InoDiag was 14.3, 23.2, 14.3, 11.9 and 26.2%, respectively. InoDiag exhibited the highest sensitivity, whereas MOMP-RB showed the best specificity. Cross-reactivity was observed with C. pneumoniae using IF, MOMP-RB and InoDiag, and Parachlamydia acanthamoebae using the cHSP60 ELISA test. No cross-reactivity was observed between C. trachomatis and the other Chlamydiales (Neochlamydia hartmannellae, Waddlia chondrophila and Simkania negevensis). Given its high sensitivity, the new automated epifluorescence immunoassay from InoDiag represents an interesting alternative. The MOMP-based ELISA of R-Biopharm should be preferred for large serological studies, given the high throughput of ELISA and its excellent specificity.
Clostridium difficile is not associated with outbreaks of viral gastroenteritis in the elderly in the NetherlandsSvraka, S.; Kuijper, E.; Duizer, E.; Bakker, D.; Koopmans, M.
doi: 10.1007/s10096-010-0913-3pmid: 20339889
The coincidental increase in norovirus outbreaks and Clostridium difficile infection (CDI) raised the question of whether these events could be related, e.g. by enhancing spread by diarrhoeal disease outbreaks. Therefore, we studied the prevalence of C. difficile in outbreaks of viral gastroenteritis in nursing homes for the elderly and characterised enzyme immunoassay (EIA)-positive stool samples. Stool samples from nursing home residents (n = 752) in 137 outbreaks of viral aetiology were investigated by EIA for the presence of C. difficile toxins. Positive samples were further tested by a cell neutralisation cytotoxicity test, a second EIA and culture. Cultured isolates were tested for the presence of toxin genes, the production of toxins and characterised by 16S rRNA polymerase chain reaction (PCR) and sequencing. Twenty-four samples (3.2%) tested positive in the EIA. Of these 24 positive samples, only two were positive by cytotoxicity and three by a second EIA. Bacterial culture of 21 available stool samples yielded a toxinogenic C. difficile PCR ribotype 001 in one patient sample only. In conclusion, we found no evidence in this retrospective study for an association between viral gastroenteritis outbreaks and C. difficile. The high rate of false-positive EIA samples emphasises the need for second confirmation tests to diagnose CDI.
Performance of rapid influenza testing in hospitalized childrenStripeli, F.; Sakkou, Z.; Papadopoulos, N.; Georgiou, V.; Gratsia, P.; Christodoulou, I.; Tsolia, M.
doi: 10.1007/s10096-010-0914-2pmid: 20349200
Influenza infection is associated with high hospitalization rates among young children. Rapid diagnosis of influenza infection is particularly useful in order to prevent nosocomial infection and allows for the timely initiation of antiviral treatment. We evaluated the performance of a rapid influenza test in hospitalized children during the influenza season. All children (aged 6 months to 14 years) hospitalized with fever and/or respiratory symptoms, admitted during the 2005 influenza season, participated in the study. A multiplex reverse transcriptase polymerase chain reaction (RT-PCR), able to identify IFV-A H1N1, H3N2, and IFV-B subtypes, was performed on nasopharyngeal aspirates. The nasal swab was tested with a lateral-flow immunoassay (QuickVue Influenza Test). The performance of the rapid test was compared with the results of PCR. Influenza infection was diagnosed by PCR in 41/217 (19%) patients. Infection with influenza A virus (H3N2) was diagnosed in all cases. The performance of the QuickVue Influenza Test was estimated as follows: sensitivity 67.5%, specificity 96%, positive predictive value 79%, and negative predictive value 93%. The sensitivity of the test was higher in infants aged 6–12 months, in those with short duration of symptoms, and in the peak phase of the epidemic. The QuickVue Influenza Test is useful and reasonably accurate to detect influenza infection in hospitalized children during the influenza season. Infection with influenza virus is unlikely if the test is negative. A positive result suggests that infection is probable if influenza virus circulates in the community.