Biomedical subjects

M C Enright

Publications and source records attributed to M C Enright.

28 records · Page 2Linked to original sources

PubMed1999

The three major Spanish clones of penicillin-resistant Streptococcus pneumoniae are the most common clones recovered in recent cases of meningitis in Spain.

One hundred six isolates of Streptococcus pneumoniae recovered in Spain from patients with meningitis in 1997 and 1998 were characterized by multilocus sequence typing. A heterogeneous collection of genotypes was associated with meningitis in Spain: 65 different sequence types were resolved and, even at a genetic distance of 0.43, there were 37 distinct lineages. Thirty-eight percent of the isolates, including all isolates of serotypes 6B, 9V, 14, and 23F, were resistant to penicillin, and 24% of the isolates were members of the three major Spanish penicillin-resistant or multidrug-resistant clones of serotypes 6B, 9V, and 23F or serotype variants of these clones. These three clones (MICs, 1 to 2 microg of penicillin/ml) were the most common clones associated with pneumococcal meningitis in Spain during 1997 and 1998. Only two of the other clones associated with meningitis were penicillin resistant (MICs, 0.12 to 0.5 microg/ml). One of the two most prevalent penicillin-susceptible clones causing meningitis (serotype 3) has not been detected outside of Spain, whereas the other (serotype 18C) has been recovered from patients with meningitis in the United Kingdom, The Netherlands, and Denmark. The prevalence of meningitis caused by isolates of the three major Spanish penicillin-resistant or multiply antibiotic-resistant clones, which are now globally distributed, is disturbing and clearly establishes their ability to cause life-threatening disease.

Base Sequence↗

PubMed1998

Recombinational exchanges at the capsular polysaccharide biosynthetic locus lead to frequent serotype changes among natural isolates of Streptococcus pneumoniae.

Serotype 19F variants of the major Spanish multiresistant serotype 23F clone of Streptococcus pneumoniae have been proposed to have arisen by recombinational exchanges at the capsular biosynthetic locus. Members of the Spanish multiresistant serotype 23F clone and the serotype 19F variants were confirmed to be essentially identical in overall genotype, as they were indistinguishable by REP-PCR, and had identical sequences at three polymorphic housekeeping genes. Eight serotype 19F variants were studied and all had large recombinational replacements at the capsular biosynthetic locus. In all cases, one of the recombinational cross-over points appeared to be upstream of dexB, which flanks one end of the capsular locus, and in six of the variants the other cross-over point was downstream of aliA, which flanks the other end of the locus. In two strains a recombinational cross-over point between the introduced serotype 19F capsular region and that of the Spanish serotype 23F clone could be clearly identified, within cpsN in one strain and within cpsM in the other. The differences in the recombinational junctions and sequence polymorphisms within the introduced capsular genes, suggested that the eight serotype 19F variants emerged on at least four separate occasions. Changes in capsular type by recombination may therefore be relatively frequent in pneumococci and this has implications for the long-term efficacy of conjugate pneumococcal vaccines that will protect against only a limited number of serotypes.

Base Sequence↗

PubMed1998

Serotype 19A variants of the Spanish serotype 23F multiresistant clone of Streptococcus pneumoniae.

Multiply-antibiotic-resistant isolates of serogroup 19 Streptococcus pneumoniae, possessing altered penicillin-binding protein (PBP) 1A, 2B, and 2X genes that are indistinguishable from those of the Spanish multiresistant serogroup 23F clone, are now commonly encountered in Spain. Those isolates that have been serotyped express type 19F capsular polysaccharide. Serotyping of further isolates, and hybridization using a serotype 19F-specific probe, has shown that some of them are serotype 19A, rather than 19F. The Spanish multiresistant serotype 19A, 19F, and 23F multiresistant strains were all shown to be very closely related in overall genotype, as they were indistinguishable by REP-PCR and by the sequencing of internal fragments of three house-keeping genes. The serotype 19A multiresistant strains, like the serotype 19F multiresistant strains, therefore appear to be a serotype variant of the Spanish multiresistant serotype 23F clone, which presumably has arisen by recombination at the capsular locus.

Bacterial Capsules↗

PubMed1998

Identification of three major clones of multiply antibiotic-resistant Streptococcus pneumoniae in Taiwanese hospitals by multilocus sequence typing.

In this paper we demonstrate the advantages of a new molecular typing procedure, multilocus sequence typing, for the unambiguous characterization of penicillin-resistant pneumococci. The sequences of approximately 450-bp fragments of seven housekeeping genes were determined for 74 penicillin-resistant Taiwanese isolates of Streptococcus pneumoniae (MIC of penicillin > 0.5 microgram/ml). The combination of alleles at the seven loci defined an allelic profile for each strain, and a dendrogram, based on the pairwise mismatches in allelic profiles, grouped 86% of the isolates into one of three penicillin-resistant clones for which the MICs of penicillin were 1 to 2 microgram/ml. Isolates within each clone had identical alleles at all seven loci or differed at only a single locus, and the fingerprints of their pbp1A, pbp2B, and pbp2X genes were uniform. Isolates of the Taiwan-19F clone and the Taiwan-23F clone were resistant to penicillin, tetracycline, and erythromycin but were susceptible to chloramphenicol. A second serotype 23F clone and serotype 19F variants of this clone were resistant to penicillin, tetracycline, chloramphenicol, and, in some cases, erythromycin. Comparisons of the allelic profiles of the three major clones with those of reference isolates of the known penicillin-resistant clones showed that the Taiwan-19F and Taiwan-23F clones were previously undescribed, whereas the second serotype 23F clone was indistinguishable from the Spanish multidrug-resistant serotype 23F clone. Single isolates of the Spanish penicillin-resistant serotype 9V clone and the Spanish multidrug-resistant serotype 6B clone were also identified in the collection.

Bacterial Typing Techniques↗

PubMed1997

Moraxella (Branhamella) catarrhalis--clinical and molecular aspects of a rediscovered pathogen.

Since its discovery at the end of the nineteenth century, Moraxella (Branhamella) catarrhalis has undergone several changes of nomenclature and periodic changes in its perceived status as either a commensal or a pathogen. Molecular analysis based on DNA hybridisation or 16S rDNA sequence comparisons has established its phylogenetic position as a member of the Moraxellaceae and shown that it is related more closely to Acinetobacter spp. than to the genus Neisseria in which it was placed formerly. However, confusion with phenotypically similar Neisseria spp. can occur in the routine diagnostic laboratory if appropriate identification tests are not performed. M. catarrhalis is now accepted as the third commonest pathogen of the respiratory tract after Streptococcus pneumoniae and Haemophilus influenzae. It is a significant cause of otitis media and sinusitis in children and of lower respiratory tract infections in adults, especially those with underlying chest disease. Nosocomial spread of infection, especially within respiratory wards, has been reported. Invasive infection is uncommon, but analysis of reports for England and Wales between 1992 and 1995 revealed 89 cases of M. catarrhalis bacteraemia, with the peak incidence in children aged 1-2 years. Carriage rates of M. catarrhalis are high in children and in the elderly, but its role as a commensal organism has probably been overstated in the past. Approximately 90% of strains are now beta-lactamase positive and, given that the first such strain was reported in 1976, this represents a dramatic increase in frequency over the last 20 years which has not been paralleled in any other species. The BRO-1 and BRO-2 beta-lactamase enzymes of M. catarrhalis are found in other Moraxellaceae, but are not related to beta-lactamases of any other species and their origin is therefore unknown. Molecular and typing studies have shown that the M. catarrhalis species is genetically heterogeneous and these methods have aided epidemiological investigation. Studies of factors that may be related to pathogenicity have shown the existence of three serotypes of lipooligosaccharide and the presence of fimbriae and a possible capsule. Some strains are serum-resistant, probably by virtue of interference with complement action, whilst transferrin- and lactoferrin-binding proteins enable the organism to obtain iron from its environment. An antibody response in humans to various M. catarrhalis antigens, including highly conserved outer-membrane proteins, has been demonstrated. Increased understanding of the organism's pathogenic properties and the host response to it may help to identify suitable vaccine targets or lead to other strategies to prevent infection. Whilst it remains, at present, the third most important respiratory pathogen, the impact of immunisation strategies for other organisms may change this position. The speed with which M. catarrhalis acquired beta-lactamase demonstrates the capacity of this organism to surprise us.

Adolescent↗

PubMed1994

Phylogenetic relationships between some members of the genera Neisseria, Acinetobacter, Moraxella, and Kingella based on partial 16S ribosomal DNA sequence analysis.

We obtained 16S ribosomal DNA (rDNA) sequence data for strains belonging to 11 species of Proteobacteria, including the type strains of Kingella kingae, Neisseria lactamica, Neisseria meningitidis, Moraxella lacunata subsp. lacunata, [Neisseria] ovis, Moraxella catarrhalis, Moraxella osloensis, [Moraxella] phenylpyruvica, and Acinetobacter lwoffii, as well as strains of Neisseria subflava and Acinetobacter calcoaceticus. The data in a distance matrix constructed by comparing the sequences supported the proposal that the genera Acinetobacter and Moraxella and [N.] ovis should be excluded from the family Neisseriaceae. Our results are consistent with hybridization data which suggest that these excluded taxa should be part of a new family, the Moraxellaceae. The strains that we studied can be divided into the following five groups: (i) M. lacunata subsp. lacunata, [N.] ovis, and M. catarrhalis; (ii) M. osloensis; (iii) [M.] phenylpyruvica; (iv) A. calcoaceticus and A. lwoffii; and (v) N. meningitidis, N. subflava, N. lactamica, and K. kingae. We agree with the previous proposal that [N.] ovis should be renamed Moraxella ovis, as this organism is closely related to Moraxella species and not to Neisseria species. The generically misnamed taxon [M.] phenylpyruvica belongs to the proposed family Moraxellaceae, but it is sufficiently different to warrant exclusion from the genus Moraxella. Further work needs to be done to investigate genetically similar species, such as Psychrobacter immobilis, before the true generic position of this organism can be determined. Automated 16S rDNA sequencing with the PCR allows workers to accurately determine phylogenetic relationships between groups of organisms.(ABSTRACT TRUNCATED AT 250 WORDS)

Acinetobacter↗

PubMed1993

Outbreak of Moraxella catarrhalis in a respiratory unit.

The clinical and epidemiological features of an outbreak of nosocomial Moraxella catarrhalis infection in a respiratory unit are described. Six isolates from five patients were shown to be indistinguishable by immunoblotting and restriction endonuclease analysis and different from 11 other, unrelated clinical strains.

Cross Infection↗

PubMed1992

Use of molecular methods to characterize Moraxella catarrhalis strains in a suspected outbreak of nosocomial infection.

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of whole cell protein, immunoblotting with normal human serum and restriction endonuclease analysis using Taq I enzyme were applied to 38 clinically significant isolates of Moraxella (Branhamella) catarrhalis obtained during a suspected outbreak of nosocomial infection. Each of 18 strains had individual profiles by at least two of the three methods (unique strains). The remaining 20 strains were assigned to five groups (A-E) on the basis of similarity by at least two of the three methods. Isolates within groups A, D and E were homologous by all three methods. Immunoblot groups B and C had two distinct whole cell protein profiles (B1 and B2) but indistinguishable restriction endonuclease profiles (group B/C). This emphasizes the need to use more than one technique in characterizing strains from suspected outbreaks of nosocomial infection. Grouped strains were more likely to originate from the same hospital ward than unique strains and were associated with a significantly longer median time from patient admission to strain isolation (14 versus 3.5 days, p less than 0.005). Furthermore, the beta-lactamase activity was homologous within the groups. The results suggest that nosocomial infection involving several distinct Moraxella catarrhalis strains persisted over a period of months, involving at least 20 patients on three different wards. Such infection is probably common in wards harbouring suitably predisposed patients. The mode of transmission remains to be elucidated, but the above three techniques possess sufficient reproducibility and discriminatory ability to constitute suitable investigative tools.

Aged↗

PubMed1992

Characterisation of hospital isolates of Moraxella (Branhamella) catarrhalis by SDS-PAGE of whole-cell proteins, immunoblotting and restriction-endonuclease analysis.

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins (WCP), immunoblot analysis and DNA restriction-endonuclease analysis (REA) were applied as potential typing methods to 31 clinically significant strains of Moraxella (Branhamella) catarrhalis, five of which came from a suspected outbreak of nosocomial infection in a respiratory-diseases ward. Twelve of 31 isolates were placed in four groups, each of which contained strains indistinguishable by the three typing techniques used. Each of a further two groups contained two strains, and they were similar by at least one technique; the remaining 15 strains were unique by all three methods. Four of five strains from the suspected outbreak were indistinguishable by SDS-PAGE of WCP, immunoblotting and REA. Results show that SDS-PAGE of WCP, immunoblotting and REA are suitable techniques for characterising M. catarrhalis and that there is a considerable degree of strain heterogeneity. Nosocomial infection with M. catarrhalis may be relatively common and further epidemiological studies with a combination of typing techniques are indicated.

Aged↗

PubMedDate not supplied

Estimating the relative contributions of mutation and recombination to clonal diversification: a comparison between Neisseria meningitidis and Streptococcus pneumoniae.

Both Neisseria meningitidis and Streptococcus pneumoniae are naturally transformable species and are known to be freely recombining in the wild. Large multilocus sequence typing (MLST) datasets have been generated for these species. Here we outline an approach which exploits these data sets in order to quantify the extent of recombination, thus enabling meaningful comparisons between the two species. Two parameters are estimated; the rate at which recombination changes alleles, compared to point mutation, and the rate at which recombination changes individual nucleotide sites, compared to point mutation. Estimates for the former parameter are 4:1 in the meningococcus (i.e. alleles are changed four-fold more frequently by recombination than by mutation), and 10:1 in the pneumococcus. However, estimates for the latter parameter are at least 80:1 in the meningococcus (i.e. an individual nucleotide site is at least 80-fold more likely to change by recombination than by mutation) and 50:1 in the pneumococcus. These data imply that recombination events, compared to mutational events, may be more common in the pneumococcus than in the meningococcus. However, because it is a more diverse species, each recombinational exchange in the meningococcus results in more nucleotide changes on average.

Genetic Variation↗