PubMed Health⌕ Search

Biomedical subjects

Michel Delmée

Publications and source records attributed to Michel Delmée.

18 recordsLinked to original sources

Third Belgian multicentre survey of antibiotic susceptibility of anaerobic bacteria.

OBJECTIVES: To collect recent data on the susceptibility of anaerobes and to compare them with results from previous studies. METHODS: Four hundred and forty-three anaerobic clinical isolates from various body sites were prospectively collected from October 2003 to February 2005 in nine Belgian hospitals. MICs were determined for nine anti-anaerobic and three recently developed antibiotics. RESULTS: Most gram-negative bacilli except Fusobacterium spp. were resistant to penicillin. Piperacillin/tazobactam, metronidazole, chloramphenicol, meropenem and amoxicillin/clavulanic acid were very active against all groups, but only 86% of Bacteroides fragilis group strains were susceptible to the latter. Cefoxitin, cefotetan and clindamycin were less active. In particular, only 62%, 52% and 48% of B. fragilis group strains were susceptible, respectively. Clindamycin shows a continuing decrease in activity, as 83% were still susceptible in 1987 and 66% in 1993-94. Anti-anaerobic activity of the new antibiotics is interesting, with MIC50 and MIC90 of 1 and >32 mg/L for moxifloxacin, 2 and 4 mg/L for linezolid and 0.5 and 8 mg/L for tigecycline. CONCLUSIONS: The susceptibility of anaerobic bacteria remains stable in Belgium, except for clindamycin, which shows a continuous decrease in activity. However, for each of the tested antibiotics, at least a few resistant organisms were detected. Consequently, for severe infections involving anaerobic bacteria, it could be advisable to perform microbiological testing instead of relying on known susceptibility profiles. Periodically monitoring background susceptibility remains necessary to guide empirical therapy.

Amoxicillin-Potassium Clavulanate Combination↗

Immunological properties of surface proteins of Clostridium difficile.

Sera from patients with Clostridium difficile-associated disease (CDAD) and sera from a control group were analysed by an ELISA to detect antibodies directed against four surface proteins and toxins A and B of C. difficile. The surface proteins were the flagellar cap protein FliD, the flagellin FliC, the adhesin Cwp66 divided into two domains, Cwp66-Nterminal and Cwp66-Cterminal, and the fibronectin-binding protein Fbp68. For each antigen, antibody levels in the CDAD patient group and in the control group were compared. In the CDAD patient group, the mean of the antibody levels decreased from Cwp66-Cterminal to Fbp68, FliD, toxins B and A, Cwp66-Nterminal and finally FliC, suggesting different immunogenic properties among these adhesins. For Cwp66-Nterminal, FliC, FliD and Fbp68, the antibody level observed in the control group was higher than in the CDAD group with a statistically significant difference whereas the antibody level for toxins A and B was not statistically different. In conclusion, this study suggests that during the clinical course of disease, C. difficile adhesins are able to induce an immune response which could play a role in the defence mechanism of the host.

Adhesins, Bacterial↗

Laboratory diagnosis of Clostridium difficile-associated diarrhoea: a plea for culture.

A routine protocol for diagnosing Clostridium difficile-associated diarrhoea (CDAD) based on both faecal-cytotoxin detection and toxigenic culture was adopted by the microbiology laboratory of the St Luc-UCL University Hospital in Brussels in 1997. A toxigenic culture is a faecal culture followed, in the case of positivity, by a direct immunoassay on colonies to detect toxin A production. The results obtained over the past 7 years in the hospital are reviewed here. A total of 10,552 diarrhoeal stools from 7042 patients were analysed, of which 9494 were negative for all tests. A total of 1058 samples (10 %) from 794 patients were culture-positive, of which 460 (4.4 %) were positive for a faecal cytotoxin. The remaining 598 cultures were tested for toxin A on colonies; 355 of them were positive, which is 3.4 % of the total, and the remaining 243 (2.3 %) were negative. The positivity of the faecal-cytotoxin assay was statistically linked to the number of colonies observed on the culture plate. In conclusion, over a 7 year period, toxigenic culture allowed the diagnosis of 355 cases of CDAD that would have been missed by a protocol using a faecal-cytotoxin assay alone. In terms of both patient care, prevention of environmental contamination and prevention of risk of a hospital outbreak, it is proposed that these results justify the recommendation to perform both faecal-toxin assay and culture in routine medical practice.

Bacteriological Techniques↗

Distribution of nocardia species in clinical samples and their routine rapid identification in the laboratory.

Eighty-six Nocardia strains isolated from clinical samples in Belgium were identified by 16S rRNA gene sequencing. Eighty-three (96%) strains belonged to only six Nocardia species: N. farcinica (38 [44%]), N. nova (19 [22%]), N. cyriacigeorgica (13 [15%]), N. brasiliensis (6 [6.9%]), N. abscessus (5 [5.8%]), and N. paucivorans (2 [2.3%]). A gallery of nine conventional and enzymatic tests was developed for the rapid identification of the most common species isolated during this survey. Pyrrolidonyl aminopeptidase, gamma-glutamyl aminopeptidase, alpha-mannosidase, and alpha-glucosidase were found to be highly discriminating and could be used to develop an identification scheme.

Algorithms↗

Histidine decarboxylase in Enterobacteriaceae revisited.

With a modification of Taylor's decarboxylation broth, histidine decarboxylase was detected in Enterobacter aerogenes, Morganella morganii, Raoultella ornithinolytica, and some strains of Citrobacter youngae and Raoultella planticola. This method provides a useful confirmatory test for identification of E. aerogenes strains.

Bacterial Typing Techniques↗

Study of genotypes and virB4 secretion gene of Bartonella henselae strains from patients with clinically defined cat scratch disease.

Bartonella henselae is the causative agent of cat scratch disease (CSD), which usually presents as a self-limiting lymphadenopathy. Occasionally, the bacteria will spread and be responsible for tissue and visceral involvement. Two B. henselae genotypes (genotypes I and II) have been described to be responsible for uncomplicated CSD on the basis of 16S rRNA sequence analysis. A type IV secretion system (T4SS) similar to the virulence-associated VirB system of Agrobacterium tumefaciens was recently identified in the B. henselae Houston-1 genotype I strain. We studied the correlations of the B. henselae genotypes with the clinical presentations and with the presence of T4SS. Isolates originated from CSD patients whose lymph nodes were prospectively analyzed. B. henselae genotype I was identified in 13 of 42 patients (30%). Among these, two teenage twins presented with hepatosplenic CSD and one immunocompetent adult presented with osteomyelitis. Genotype II was detected in 28 of 42 patients (67%), all of whom presented with uncomplicated CSD. The last patient was infected with both genotypes. T4SS was studied by PCR amplification of the virB4 gene. Amplification of virB4 codons 146 to 256, 273 to 357, and 480 to 537 enabled us to detect 66, 90, and 100% of the B. henselae isolates, respectively. Sequence analysis revealed sequence variations that correlated with genotype distribution. Our studies suggest that B. henselae genotype I strains harbor virB4 genes that are different from those harbored by genotype II strains and that genotype I strains might be more pathogenic.

Adolescent↗

Evaluation of Oxoid combination discs for detection of extended-spectrum beta-lactamases.

OBJECTIVES: We evaluated the reliability of cefpirome/clavulanate (CD04) compared with ceftazidime/clavulanate (CD02) and cefotaxime/clavulanate (CD03) Oxoid combination discs for the detection of extended-spectrum beta-lactamases (ESBL) in several Enterobacteriaceae isolates, including Enterobacter spp. METHODS: Overall, a total of 105 ESBL-positive [positive double-disc synergy test (DDST)] and 94 ESBL-negative (negative DDST) Gram-negative isolates were evaluated. Ninety-eight isolates were confirmed as ESBL-positive on the basis of the sequence alignments of the blaTEM and/or blaSHV gene amplification products, which matched with previously identified ESBLs. The phenotypic detection of ESBLs was performed by the three combination discs according to the NCCLS and BSAC methods. The CD04 disc was evaluated with the manufacturer's recommended zone size difference breakpoint of > or =4 mm. RESULTS: In Escherichia coli and Klebsiella spp., the sensitivities (%)/specificities (%) of CD02, CD03 and CD04 discs, and the combination of CD02 or CD04 discs, were, respectively, 88/92, 90/92, 95/84 and 100/82, while the corresponding figures were 94/100, 4/100, 94/100 and 100/100 in Enterobacter aerogenes. NCCLS and BSAC methods yielded concordant results in 99% of the isolates. CONCLUSIONS: CD04 and CD02 discs were the best combination for detection of ESBLs in our collection of Enterobacteriaceae isolates, including E. aerogenes.

Cephalosporins↗

Infection by Mycoplasma hyorhinis strongly enhances uptake of antisense oligonucleotides: a reassessment of receptor-mediated endocytosis in the HepG2 cell line.

This paper shows that the approximately 66 kDa band, previously isolated from the HepG2 cell line as an oligonucleotide (ON) plasma membrane 'receptor', is induced by Mycoplasma infection. Moreover, this band has been identified as the invariant membrane protein of Mycoplasma hyorhinis, p70, based on ribosomal DNA sequencing combined with ON ligand blotting after p70 immunoprecipitation by a monoclonal antibody. Whereas antibiotic treatment of infected HepG2 cells strongly decreased ON capture, as measured by a biochemical assay, conversely, deliberate infection of HeLa cells with M.hyorhinis dramatically promoted ON uptake but did not affect receptor-mediated endocytosis of transferrin. This was confirmed by confocal microscopy of infected HepG2 cells, which also showed an indistinguishable labelling pattern after exposure of living cells to fluorescent ON and after p70 immunolabelling in permeabilised fixed cells. We propose that ON binds to p70 on M.hyorhinis attached at the cell surface, after which the complex is internalised by 'piggy-back' endocytosis.

Anti-Bacterial Agents↗

Brevibacterium lutescens sp. nov., from human and environmental samples.

Three strains of coryneform rods isolated from clinical samples and one of environmental origin exhibited phenotypic and chemotaxonomic properties characteristic of the genus Brevibacterium and their 16S rRNA gene sequences were closely related (98.5-99.0 %) to that of Brevibacterium otitidis. However, DNA-DNA hybridization of one strain (CF87(T)) showed only 59.6 % relatedness to the type strain of B. otitidis, DSM 10718(T), and 75-82 % relatedness to the three other strains. The four strains could be differentiated from B. otitidis by cellular fatty acid composition and some phenotypic characteristics. These findings suggest that the four strains belong to a novel species, for which the name Brevibacterium lutescens sp. nov. is proposed. The type strain of B. lutescens is CF87(T) (=DSM 15022(T)=CCUG 46604(T)).

Brevibacterium↗

International typing study of toxin A-negative, toxin B-positive Clostridium difficile variants.

Clinically important strains of Clostridium difficile that do not produce toxin A but produce toxin B and are cytotoxic (A(-)/B(+)) have been reported from multiple countries. In order to compare the relatedness of these strains, we typed 23 A(-)/B(+) C. difficile isolates from the United Kingdom (6 isolates), Belgium (11 isolates), and the United States (6 isolates) by three well-described typing methods. Restriction endonuclease analysis (REA), PCR ribotyping, and serogrouping differentiated 11, 4, and 3 different strain types, respectively. Twenty-one of the 23 A(-)/B(+) variants had a 1.8-kb truncation of the toxin A gene characteristic of toxinotype VIII strains; 20 of the 21 toxinotype VIII-like strains were PCR type 17. PCR type 17 isolates could be differentiated into two separate strain groups by serogrouping and by REA. REA further discriminated these isolates into eight subgroups (REA types). PCR type 17-serogroup F-REA group CF isolates were recovered from all three countries, and one specific REA type, CF4, was recovered from patients with C. difficile disease in the United Kingdom and the United States. C. difficile A(-)/B(+) variants of apparent clonal origin are widely distributed in Europe and North America.

Adult↗

Bacteremia due to a novel Microbacterium species in a patient with leukemia and description of Microbacterium paraoxydans sp. nov.

A yellow-pigmented coryneform rod was isolated from the blood of a child with acute lymphoblastic leukemia who was perfused with a central venous catheter. The culture bottles were positive twice, at a 2-month interval. The isolate was identified as a Microbacterium sp. and studied along with five other similar strains. Phenotypic, chemotaxonomic, and genetic characteristics indicated that they are closely related to Microbacterium oxydans but that they belong to a distinct species, for which the name Microbacterium paraoxydans sp. nov. is proposed. The type strain of M. paraoxydans is CF36(T) = DSM 15019(T). The G+C content of its DNA is 69.9 mol%.

Actinomycetales↗

Osteomyelitis caused by Bartonella henselae genotype I in an immunocompetent adult woman.

We report the first description of osteomyelitis due to Bartonella henselae genotype I in an immunocompetent middle-aged woman. The diagnosis was established by serology, histopathology, and PCR analysis of osseous and lymph node tissues. The mycobacteria growth indicator tube inoculated with the lymph node aspirate was used for PCR analysis.

Antibodies, Bacterial↗

Biochemical and susceptibility tests useful for identification of nonfermenting gram-negative rods.

Six hundred nineteen strains of nonfermenting gram-negative rods were tested for alkaline phosphatase, benzyl-arginine arylamidase, pyrrolidonyl arylamidase, ethylene glycol acidification, and susceptibility to desferrioxamine and colistin. The results were highly discriminant. Therefore, the proposed tests may be helpful for the identification of this group of organisms.

Alkaline Phosphatase↗

Clostridium difficile genotyping based on slpA variable region in S-layer gene sequence: an alternative to serotyping.

Recent investigations of Clostridium difficile cell wall components have revealed the presence of an S-layer encoded by the slpA gene. The aim of this study was to determine whether slpA genotyping can be used as an alternative to serotyping. The variable regions of slpA were amplified by PCR from serogroup reference strains and various clinical isolates chosen randomly. Amplified products were analyzed after restriction enzyme digestion and DNA sequencing. The sequences of the variable region of the SlpA protein were found to be strictly identical within a given serogroup but divergent between serogroups. These preliminary results suggest that PCR-restriction fragment length polymorphism, in conjunction with DNA sequencing of the slpA variable region, could constitute an alternative typing method for determining C. difficile serotypes.

Amino Acid Sequence↗

Toxin A detection on Clostridium difficile colonies from 24-h cultures.

OBJECTIVE: Performance of a combined approach for the detection of toxigenic strains in patients suspected of having Clostridium difficile-associated disease was evaluated. METHODS: In this approach, stools were cultured for 24 h on a selective medium supplemented with sodium taurocholate (TCCFA), in anaerobic conditions created with the Martreg Anoxomat system, and toxin A detection was performed directly on C. difficile colonies, by enzyme immunoassay (EIA). This method was compared with three others: cytotoxigenic culture consisting of a 48-h culture on selective medium followed by detection of in vitro cytotoxin production on cell monolayers, fecal cytotoxin detection and fecal toxin A detection by EIA. RESULTS: From 548 stools, 108 yielded a positive culture by at least one of the methods, and 81 isolates were cytotoxin producers. Cultures for 24 h on TCCFA were positive in 106 cases and EIA performed on colonies gave 73 positive results, giving a sensitivity of 90.1% and a specificity of 100%. By comparison, the sensitivity and specificity of cytotoxigenic culture, stool cytotoxin and stool toxin A were respectively 96.2% and 100%, 61.7% and 100%, and 66.7% and 95.9%. CONCLUSIONS: Performing EIA on colonies recovered after 24 h culture allows us to improve the detection of toxigenic strains in patients suspected of having C. difficile-associated disease.

Journal Article↗