Fingerprinting Acinetobacter strains from clinical sources by numerical analysis of electrophoretic protein patterns

Alexander, M.; Ismail, Farzana; Jackman, P. J. H.; Noble, W. C.

doi:10.1099/00222615-18-1-55

Cited by 63 publications

(28 citation statements)

References 12 publications

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“…Typing by PAGE overcomes such problems especially as the same approach can be used for other organisms (which for serotyping would each require a large range of sera). Other organisms to which PAGE typing has been applied include Acinetobacter (Alexander et al 1984), Bacteroides (Taylor et al 1987), Campylobacter (Costas et al 1987b), Clostridium (Tabaqchali et al 1986) and Streptobacillus .…”

Section: Discussionmentioning

confidence: 99%

Numerical analysis of electrophoretic protein patterns of Providencia rettgeri strains from human faeces, urine and other specimens

Costas¹,

Holmes²,

Wood³

et al. 1989

Journal of Applied Bacteriology

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Thirty-one strains of Providencia rettgeri (mainly from humans) were characterized by one-dimensional SDS-PAGE of cellular proteins. The strains came from various countries and comprised 14 from urine, eight from faeces, two from bile (plus one from the liver of a sheep), two from sputum, one from an insect pupa and three the sources of which were unknown. Also included, for reference purposes, were the type strains of the four other Providencia species. The protein patterns, which contained 45-50 discrete bands, were highly reproducible and were used as the basis for two numerical analyses. In the first, which included all the protein bands, the 31 Prov. rettgeri strains formed 13 clusters at the 84% S level. In the second analysis, in which the principal protein bands (in the 33.3-41.3 kD range) were excluded, 29 of the 31 Prov. rettgeri strains formed a single cluster at the 81% S level, whilst the four Providencia reference strains remained unclustered. We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides the basis for typing clinical strains of Prov. rettgeri. Reference strains of each of the 13 PAGE types identified are available from NCTC for inclusion in future studies.

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Section: Discussionmentioning

confidence: 99%

Numerical analysis of electrophoretic protein patterns of Providencia rettgeri strains from human faeces, urine and other specimens

Costas¹,

Holmes²,

Wood³

et al. 1989

Journal of Applied Bacteriology

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“…(1956). The protein content of sludge sample was determined by resuspending harvested sludge an equal volume of sample treatment buffer (0·0625 M Tris HCl buffer, pH 6·8, sodium dodecyl sulphate (SDS) 2%, glycerol 10%, 2‐mercaptoethanol 5%), and was heated at 100°C for 10 min (Alexander et al . 1984).…”

Section: Methodsmentioning

confidence: 99%

The role of cellular polysaccharides in the formation and stability of aerobic granules

Tay

Liu

2001

Lett Appl Microbiol

253

138

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Aims: This paper attempts to investigate the role of cellular polysaccharides in the formation and stability of aerobic granules. Methods and Results: Three column sequential aerobic sludge blanket reactors (R1, R2 and R3) were operated at a super®cial air up¯ow velocity of 0á3 cm s ±1 , 1á2 cm s ±1 and 2á4 cm s ±1 , respectively. Aerobic granules appeared at cycle 42 in R2 and R3 with a mean size of 0á37 mm in R2 and 0á35 mm in R3, however, aerobic granulation was not observed in R1. After the formation of aerobic granules, the sludge volume index (SVI) decreased to 55 ml g ±1 in R2 and 46 ml g ±1 in R3. Aerobic granulation was concurrent with a sharp increase of cellular polysaccharides normalized to cellular proteins, which increased from 5á7 to 13á0 mg per mg proteins in R2, and 7á5±13á9 mg per mg protein in R3. The content of polysaccharides in aerobic granules was 2±3 times higher than that in the bio¯occi cultivated in R1. The disappearance of aerobic granules in R2 was tightly coupled to a drop in cellular polysaccharides. After the reappearance of bio¯occi in R2, the content of cellular polysaccharides were found to be restored to the level observed in R1.Conclusions: It appears that the production of cellular polysaccharides could be stimulated by hydrodynamic shear force and contributes to the formation and stability of aerobic granules. Signi®cance and Impact of the Study: It is expected that this study would provide useful information for better understanding the mechanisms of aerobic granulation.

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“…A number of recent studies have attempted to set up reliable identification schemes based on DNA homology, phenotypic characters, and comparison of cell envelope and outer membrane protein patterns, but such methods yield inconsistent results (1,10,11,12,23). The phenotypic characters of Acinetobacter species are susceptible to the conditions under which they are cultured (10).…”

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confidence: 99%

Sequencing of the rpoB Gene and Flanking Spacers for Molecular Identification of Acinetobacter Species

et al. 2006

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Acinetobacter species are defined on the basis of several phenotypic characters, results of DNA-DNA homology, and more recently, similarities or dissimilarities in 16S rRNA gene sequences. However, the 16S rRNA gene is not polymorphic enough to clearly distinguish all Acinetobacter species. We used an RNA polymerase ␤-subunit gene (rpoB)-based identification scheme for the delineation of species within the genus Acinetobacter, and towards that end, we determined the complete rpoB gene and flanking spacer (rplL-rpoB and rpoB-rpoC) sequences of the 17 reference strains of Acinetobacter species and 7 unnamed genomospecies. By using complete gene sequences (4,089 bp), we clearly separated all species and grouped them into different clusters. A phylogenetic tree constructed using these sequences was supported by bootstrap values higher than those obtained with 16S rRNA or the gyrB or recA gene. Four pairs of primers enabled us to amplify and sequence two highly polymorphic partial sequences (350 and 450 bp) of the rpoB gene. These and flanking spacers were designed and tested for rapid identification of the 17 reference strains of Acinetobacter species and 7 unnamed genomospecies. Each of these four variable sequences enabled us to delineate most species. Sequences of at least two polymorphic sequences should be used to distinguish Acinetobacter grimontii, Acinetobacter junii, Acinetobacter baylyi, and genomic species 9 from one another. Finally, 21 clinical isolates of Acinetobacter baumannii were tested for intraspecies relationships and assigned correctly to the same species by comparing the partial sequences of the rpoB gene and its flanking spacers.Acinetobacter species are ubiquitous in the environment and have emerged as important nosocomial pathogens (3). A propensity to tolerate drying (29, 30) and resistance to many commonly used antibiotics (28) are key factors in enabling the organism to survive and spread in the nosocomial environment. Furthermore, Acinetobacter species may serve as reservoirs of antibiotic-resistant genes, particularly in hospital environments (22).The genus Acinetobacter, originally proposed by Brisou and Prevot (6), comprises a collection of bacteria which show a great deal of phenotypic similarities. Currently, the genus Acinetobacter comprises many taxons based on hybridization groups and classified as genomovars and genomospecies. From an official taxonomic point of view, there are currently 17 Acinetobacter nomenspecies with standing in nomenclature. However, several other genomospecies have been proposed, mainly on the basis of DNA hybridization studies (7,20,21,28).Delineation of species within the genus Acinetobacter is often problematic (22). A number of recent studies have attempted to set up reliable identification schemes based on DNA homology, phenotypic characters, and comparison of cell envelope and outer membrane protein patterns, but such methods yield inconsistent results (1,10,11,12,23). The phenotypic characters of Acinetobacter species are susceptible to the condit...

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Fingerprinting Acinetobacter strains from clinical sources by numerical analysis of electrophoretic protein patterns

Cited by 63 publications

References 12 publications

Numerical analysis of electrophoretic protein patterns of Providencia rettgeri strains from human faeces, urine and other specimens

Numerical analysis of electrophoretic protein patterns of Providencia rettgeri strains from human faeces, urine and other specimens

The role of cellular polysaccharides in the formation and stability of aerobic granules

Sequencing of the rpoB Gene and Flanking Spacers for Molecular Identification of Acinetobacter Species

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