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FEMS microbiology letters2008; 284(2); 247-252; doi: 10.1111/j.1574-6968.2008.01208.x

Evaluation of a multiplex PCR for detection of serotypes K1, K2 and K5 in Klebsiella sp. and comparison of isolates within these serotypes.

Abstract: A multiplex PCR using targets within the serotype-specific region of the capsular polysaccharide synthesis gene cluster of serotypes K1, K2 and K5 was evaluated using the 77 reference serotype strains of Klebsiella, and a panel of clinical isolates subjected previously to conventional serotyping. The PCR was highly specific for these serotypes, which are those most associated with virulence in humans and horses. PCR confirmed that isolates of the K5 serotype had cross-reacted with antiserum for other serotypes, particularly for K7. K5 isolates received by our laboratory were almost exclusively from thoroughbred horses, and were submitted for screening prior to breeding programmes. Most, including a reference strain isolated in 1955, belonged to a cluster of genetically similar isolates of sequence type (ST) 60. K1 isolates, all from humans, belonged to a previously identified cluster of ST 23.
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Publication Date: 2008-05-27 PubMed ID: 18507682DOI: 10.1111/j.1574-6968.2008.01208.xGoogle Scholar: Lookup
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  • Comparative Study
  • Evaluation Study
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research evaluated the efficacy of a multiplex PCR in identifying specific serotypes (K1, K2, K5) of the bacteria Klebsiella, validating the tool using a collection of both reference and clinically isolated strains. The study also illuminated some clinical implications of these serotypes in humans and horses, and identified potential genetic clusters of similar isolates within specific serotypes.

Methodology and Findings

  • The researchers used a multiplex PCR, a method that allows multiple targets to be amplified simultaneously in one PCR experiment, as their detection tool. They selected targets within the serotype-specific region of the capsular polysaccharide synthesis gene cluster of serotypes K1, K2, and K5 of Klebsiella.
  • The effectiveness of this PCR method was evaluated using 77 reference serotype strains, acting as a control group, and a panel of clinically isolated strains that had undergone conventional serotyping.
  • The study found that the multiplex PCR was highly specific for these serotypes (K1, K2, and K5), which are the most associated with virulence, the ability to cause disease, in both humans and horses.

Clinical Implications and Observations

  • The researchers observed that some K5 serotype isolates had cross-reacted with antiserum for other serotypes, particularly K7, indicating potential misidentifications in serotyping.
  • The K5 isolates, which were mostly from thoroughbred horses, were found to be frequently submitted for screening during horse breeding programmes.

Genetic Clusters Within Serotypes

  • In their analysis, they identified specific genetic clusters, or groups of genetically similar isolates, within certain serotypes.
  • Most K5 isolates, including a reference strain isolated as far back as 1955, belonged to a cluster of isolates identified as sequence type (ST) 60.
  • In contrast, all K1 isolates, which were all sourced from human samples, belonged to ST 23, a previously identified cluster.

Cite This Article

APA
Turton JF, Baklan H, Siu LK, Kaufmann ME, Pitt TL. (2008). Evaluation of a multiplex PCR for detection of serotypes K1, K2 and K5 in Klebsiella sp. and comparison of isolates within these serotypes. FEMS Microbiol Lett, 284(2), 247-252. https://doi.org/10.1111/j.1574-6968.2008.01208.x

Publication

FEMS microbiology letters
ISSN: 0378-1097
NlmUniqueID: 7705721
Country: England
Language: English
Volume: 284
Issue: 2
Pages: 247-252

Researcher Affiliations

Turton, Jane F
  • Laboratory of HealthCare Associated Infection, Centre for Infections, Health Protection Agency, London, UK. jane.turton@hpa.org.uk
Baklan, Hatice
    Siu, L K
      Kaufmann, Mary E
        Pitt, Tyrone L

          MeSH Terms

          • Animals
          • Antigens, Bacterial
          • Bacterial Capsules / genetics
          • Cross Reactions
          • DNA, Bacterial / isolation & purification
          • Electrophoresis, Gel, Pulsed-Field
          • Genes, Bacterial
          • Horses
          • Humans
          • Klebsiella / classification
          • Klebsiella / genetics
          • Multigene Family
          • Polymerase Chain Reaction / methods
          • Polysaccharides, Bacterial
          • Sensitivity and Specificity
          • Serotyping / methods

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