Klebsiella typing: pulsed-field gel electrophoresis (PFGE) in comparison with O:K-serotyping

Hansen, Dennis Schrøder; Skov, Robert; Benedí, J.V.; Sperling, V.; Kolmos, Hans Jørn

doi:10.1046/j.1469-0691.2002.00411.x

Cited by 32 publications

(27 citation statements)

References 25 publications

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“…Strain typing is necessary to recognize clonal groups, to assist epidemiological investigations, and to link genetic diversity with pathophysiological specificities. Pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and capsular polysaccharide characterization (K typing) are widely used to characterize K. pneumoniae clinical isolates (4)(5)(6)(7). MLST has been widely adopted as the common language for the characterization of K. pneumoniae and the recognition of important emerging sequence types (STs), such as ST23, which is associated with liver abscesses (8), and ST15, which is associated with multidrug resistance worldwide (9,10).…”

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confidence: 99%

“…On the contrary, allele-specific PCRs are easy to perform but were mainly developed for serotypes that are frequent in communityacquired bacteremia and liver abscess isolates, namely, K. pneumoniae K1, K2, K5, K20, K54, and K57 (14,17,20,21). However, these serotypes represent a small proportion (Ͻ20%) of K. pneumoniae isolates from sources other than liver abscesses or community-acquired pneumonia (6,22,23), thus leaving most of the classical K. pneumoniae isolates untypeable. Although it should be feasible to develop specific PCR assays for most K types, it is not practical given the limitations of PCR multiplexing.…”

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confidence: 99%

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wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

et al. 2013

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f Pathogens of the genus Klebsiella have been classified into distinct capsular (K) types for nearly a century. K typing of Klebsiella species still has important applications in epidemiology and clinical microbiology, but the serological method has strong practical limitations. Our objective was to evaluate the sequencing of wzi, a gene conserved in all capsular types of Klebsiella pneumoniae that codes for an outer membrane protein involved in capsule attachment to the cell surface, as a simple and rapid method for the prediction of K type. The sequencing of a 447-nucleotide region of wzi distinguished the K-type reference strains with only nine exceptions. A reference wzi sequence database was created by the inclusion of multiple strains representing K types associated with high virulence and multidrug resistance. A collection of 119 prospective clinical isolates of K. pneumoniae were then analyzed in parallel by wzi sequencing and classical K typing. Whereas K typing achieved typeability for 81% and discrimination for 94.4% of the isolates, these figures were 98.1% and 98.3%, respectively, for wzi sequencing. The prediction of K type once the wzi allele was known was 94%. wzi sequencing is a rapid and simple method for the determination of the K types of most K. pneumoniae clinical isolates.

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wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

et al. 2013

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“…Following the pioneering work of Julianelle (17), who described the first three capsular serotypes, 77 K antigens are now included in the international K-typing scheme (27,28). Determination of the K type has been used since the 1920s to type Klebsiella isolates (17) and has long been the preferred method for the investigation of epidemiological relationships among strains (11,13,14,18,28). Because it is highly discriminatory and reproducible, K serotyping is particularly useful for the comparison of strains from different geographical locations and different times.…”

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confidence: 99%

“…ozaenae and K. pneumoniae subsp. rhinoscleromatis isolates, the following biochemical tests were used (14): Voges-Proskauer test and tests for urease, o-nitrophenyl-␤-D-galactopyranoside, lysine decarboxylase, citrate, malonate, and gas production.…”

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Molecular Serotyping ofKlebsiellaSpecies Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster

2004

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The objective of the present work was to develop a molecular method that would enable determination of the capsular serotypes of Klebsiella isolates without the use of antiserum. PCR amplification of the capsular antigen gene cluster (cps) was followed by digestion with the restriction enzyme HincII (cps PCR-restriction fragment length polymorphism [RFLP] analysis). The profiles (C patterns) obtained for 224 strains representing the 77 known K serotypes showed 3 to 13 fragments ranging in size from 0.2 to 4.4 kb. A total of 97 distinct C patterns were obtained; 100% of 61 pairs of samples tested twice showed reproducible C patterns. The C patterns were K-type specific; i.e., the C pattern(s) of any K serotype was distinct from the C patterns of all other K serotypes, with the only exceptions being serotypes K22 and K37, which are known to cross-react. For 12 of 17 K types for which at least two strains were included, C-pattern variations were found among strains with the same K serotype. Therefore, cps PCR-RFLP analysis has a higher discriminatory power than classical K serotyping. C-pattern identity was observed among strains with a given K type that were collected many years apart and from distinct sources, indicating C-pattern stability. Only 4.5% of the strains were nontypeable, because of unsuccessful PCR amplification (whereas 8 to 23% are nontypeable by classical K serotyping). Three of four noncapsulated strains analyzed showed recognizable C patterns. The K serotypes of 18 (82%) of 22 recent Klebsiella pneumoniae clinical isolates could be deduced from their C patterns. In conclusion, cps PCR-RFLP analysis allows determination of the K serotype, while it is easier to perform and more discriminatory than classical serotyping.

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“…El método de genotipificación bacteriana por PFGE es el recomendado en el estudio de brotes de Klebsiella, debido a su alto poder discriminatorio de los aislamientos relacionados genéticamente (27).…”

Section: Discussionunclassified

Caracterización molecular de cepas de Klebsiella pneumoniae productoras de BLEE causantes de infección intrahospitalaria en el servicio de neonatología de un hospital de Lima, Perú

Bailón

Sacsaquispe

2013

Rev Med Hered

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Klebsiella typing: pulsed-field gel electrophoresis (PFGE) in comparison with O:K-serotyping

Cited by 32 publications

References 25 publications

wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

Molecular Serotyping ofKlebsiellaSpecies Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster

Caracterización molecular de cepas de Klebsiella pneumoniae productoras de BLEE causantes de infección intrahospitalaria en el servicio de neonatología de un hospital de Lima, Perú

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