Literature DB >> 3312300

Automation in clinical microbiology: a new approach to identifying micro-organisms by automated pattern matching of proteins labelled with 35S-methionine.

S Tabaqchali1, R Silman, D Holland.   

Abstract

A new rapid automated method for the identification and classification of microorganisms is described. It is based on the incorporation of 35S-methionine into cellular proteins and subsequent separation of the radiolabelled proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The protein patterns produced were species specific and reproducible, permitting discrimination between the species. A large number of Gram negative and Gram positive aerobic and anaerobic organisms were successfully tested. Furthermore, there were sufficient differences within species between the protein profiles to permit subdivision of the species. New typing schemes for Clostridium difficile, coagulase negative staphylococci, and Staphylococcus aureus, including the methicillin resistant strains, could thus be introduced; this has provided the basis for useful epidemiological studies. To standardise and automate the procedure an automated electrophoresis system and a two dimensional scanner were developed to scan the dried gels directly. The scanner is operated by a computer which also stores and analyses the scan data. Specific histograms are produced for each bacterial species. Pattern recognition software is used to construct databases and to compare data obtained from different gels: in this way duplicate "unknowns" can be identified. Specific small areas showing differences between various histograms can also be isolated and expanded to maximise the differences, thus providing differentiation between closely related bacterial species and the identification of differences within the species to provide new typing schemes. This system should be widely applied in clinical microbiology laboratories in the near future.

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Year:  1987        PMID: 3312300      PMCID: PMC1141175          DOI: 10.1136/jcp.40.9.1070

Source DB:  PubMed          Journal:  J Clin Pathol        ISSN: 0021-9746            Impact factor:   3.411


  35 in total

1.  Identification and grouping of bacteria by numerical analysis of their electrophoretic protein patterns.

Authors:  K Kersters; J De Ley
Journal:  J Gen Microbiol       Date:  1975-04

2.  New method for typing Staphylococcus aureus resistant to methicillin based on sulphur-35 methionine labelled proteins: its application in an outbreak.

Authors:  J R Stephenson; S J Crook; S Tabaqchali
Journal:  Br Med J (Clin Res Ed)       Date:  1986-09-06

3.  Immunoblotting to demonstrate antigenic and immunogenic differences among nine standard strains of Clostridium difficile.

Authors:  S R Heard; B Rasburn; R C Matthews; S Tabaqchali
Journal:  J Clin Microbiol       Date:  1986-09       Impact factor: 5.948

4.  Quantitative comparison of electrophoretic traces of bacterial proteins.

Authors:  R K Feltham; P H Sneath
Journal:  Comput Biomed Res       Date:  1979-06

Review 5.  Nucleic acids, genetic exchange and bacterial speciation.

Authors:  S Falkow
Journal:  Am J Med       Date:  1965-11       Impact factor: 4.965

6.  Cleavage of structural proteins during the assembly of the head of bacteriophage T4.

Authors:  U K Laemmli
Journal:  Nature       Date:  1970-08-15       Impact factor: 49.962

7.  A comparison of the 16S ribosomal RNAs from mesophilic and thermophilic bacilli: some modifications in the Sanger method for RNA sequencing.

Authors:  C Woese; M Sogin; D Stahl; B J Lewis; L Bonen
Journal:  J Mol Evol       Date:  1976-04-09       Impact factor: 2.395

8.  The phylogeny of prokaryotes.

Authors:  G E Fox; E Stackebrandt; R B Hespell; J Gibson; J Maniloff; T A Dyer; R S Wolfe; W E Balch; R S Tanner; L J Magrum; L B Zablen; R Blakemore; R Gupta; L Bonen; B J Lewis; D A Stahl; K R Luehrsen; K N Chen; C R Woese
Journal:  Science       Date:  1980-07-25       Impact factor: 47.728

9.  Relationships between strains of Mycoplasma mycoides subspp. mycoides and capri studied by two-dimensional gel electrophoresis of cell proteins.

Authors:  A W Rodwell; E S Rodwell
Journal:  J Gen Microbiol       Date:  1978-12

10.  The classification of staphylococci from colonized ventriculo-atrial shunts.

Authors:  R Holt
Journal:  J Clin Pathol       Date:  1969-07       Impact factor: 3.411

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  4 in total

1.  Evaluation of fingerprinting methods for identification of Helicobacter pylori strains.

Authors:  C L Clayton; H Kleanthous; J C Dent; C A McNulty; S Tabaqchali
Journal:  Eur J Clin Microbiol Infect Dis       Date:  1991-12       Impact factor: 3.267

2.  Differentiation and characterization of enteroviruses by computer-assisted viral protein fingerprinting.

Authors:  D T Holland; J Senne; C R Peter; C Urmenta; J D Connor
Journal:  J Clin Microbiol       Date:  1998-06       Impact factor: 5.948

3.  Electrophoretic protein typing of Campylobacter jejuni subspecies "doylei" (nitrate-negative campylobacter-like organisms) from human faeces and gastric mucosa.

Authors:  R J Owen; M Costas; L L Sloss
Journal:  Eur J Epidemiol       Date:  1988-09       Impact factor: 8.082

4.  Molecular studies on the epidemiology and pathogenicity of Clostridium difficile.

Authors:  S Tabaqchali
Journal:  Gut       Date:  1989-11       Impact factor: 23.059

  4 in total

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