Identification of plasmids by PCR-based replicon typing

Alessandra Carattoli; Alessia Bertini; Laura Villa; Vincenzo Falbo; Katie L. Hopkins; E. John Threlfall

doi:10.1016/j.mimet.2005.03.018

Identification of plasmids by PCR-based replicon typing

Alessandra Carattoli^*, Alessia Bertini, Laura Villa, Vincenzo Falbo, Katie L. Hopkins, E. John Threlfall

^*Corresponding author for this work

HCAI and AMR Divisional Office

Research output: Contribution to journal › Article › peer-review

2076 Citations (Scopus)

Abstract

The epidemiological importance of tracing plasmids conferring drug resistance prompted us to develop a PCR method based on replicons (inc/rep PCR) of the major plasmid incompatibility groups among Enterobacteriaceae. Eighteen pairs of primers were designed to perform 5 multiplex- and 3 simplex-PCRs, recognizing FIA, FIB, FIC, HI1, HI2, I1-Iγ, L/M, N, P, W, T, A/C, K, B/O, X, Y, F, and FIIA. The specificity of the method was tested on a collection of 61 reference plasmids and on 20 Salmonella enterica strains of different serotypes isolated in Italy. Results indicated that the inc/rep PCR method demonstrates high specificity and sensitivity in detecting replicons on reference plasmids and also revealed the presence of recurrent and common plasmids in epidemiologically unrelated Salmonella isolates of different serotypes. These results suggest that the method is potentially applicable to a large number of strains to trace the diffusion of specific multi-drug resistance plasmids in different environments.

Original language	English
Pages (from-to)	219-228
Number of pages	10
Journal	Journal of Microbiological Methods
Volume	63
Issue number	3
DOIs	https://doi.org/10.1016/j.mimet.2005.03.018
Publication status	Published - Dec 2005

Bibliographical note

Funding Information:
We are grateful to V. Miriagou, L.S. Tzouvelekis, E. Tzelepi and G.M. Rossolini for helpful discussion and continuous encouragement. We thank Fabio Riccobono for the DNA sequencing. This work was supported by grants from the Istituto Superiore di Sanità (Art. 502, project no. 2012/RI; Art. 524, project no. C3MD).

Keywords

PCR
Plasmids
Replicon

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10.1016/j.mimet.2005.03.018

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title = "Identification of plasmids by PCR-based replicon typing",

abstract = "The epidemiological importance of tracing plasmids conferring drug resistance prompted us to develop a PCR method based on replicons (inc/rep PCR) of the major plasmid incompatibility groups among Enterobacteriaceae. Eighteen pairs of primers were designed to perform 5 multiplex- and 3 simplex-PCRs, recognizing FIA, FIB, FIC, HI1, HI2, I1-Iγ, L/M, N, P, W, T, A/C, K, B/O, X, Y, F, and FIIA. The specificity of the method was tested on a collection of 61 reference plasmids and on 20 Salmonella enterica strains of different serotypes isolated in Italy. Results indicated that the inc/rep PCR method demonstrates high specificity and sensitivity in detecting replicons on reference plasmids and also revealed the presence of recurrent and common plasmids in epidemiologically unrelated Salmonella isolates of different serotypes. These results suggest that the method is potentially applicable to a large number of strains to trace the diffusion of specific multi-drug resistance plasmids in different environments.",

keywords = "PCR, Plasmids, Replicon",

author = "Alessandra Carattoli and Alessia Bertini and Laura Villa and Vincenzo Falbo and Hopkins, {Katie L.} and Threlfall, {E. John}",

note = "Funding Information: We are grateful to V. Miriagou, L.S. Tzouvelekis, E. Tzelepi and G.M. Rossolini for helpful discussion and continuous encouragement. We thank Fabio Riccobono for the DNA sequencing. This work was supported by grants from the Istituto Superiore di Sanit{\`a} (Art. 502, project no. 2012/RI; Art. 524, project no. C3MD).",

year = "2005",

month = dec,

doi = "10.1016/j.mimet.2005.03.018",

language = "English",

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AU - Carattoli, Alessandra

AU - Bertini, Alessia

AU - Villa, Laura

AU - Falbo, Vincenzo

AU - Hopkins, Katie L.

AU - Threlfall, E. John

N1 - Funding Information: We are grateful to V. Miriagou, L.S. Tzouvelekis, E. Tzelepi and G.M. Rossolini for helpful discussion and continuous encouragement. We thank Fabio Riccobono for the DNA sequencing. This work was supported by grants from the Istituto Superiore di Sanità (Art. 502, project no. 2012/RI; Art. 524, project no. C3MD).

PY - 2005/12

Y1 - 2005/12

N2 - The epidemiological importance of tracing plasmids conferring drug resistance prompted us to develop a PCR method based on replicons (inc/rep PCR) of the major plasmid incompatibility groups among Enterobacteriaceae. Eighteen pairs of primers were designed to perform 5 multiplex- and 3 simplex-PCRs, recognizing FIA, FIB, FIC, HI1, HI2, I1-Iγ, L/M, N, P, W, T, A/C, K, B/O, X, Y, F, and FIIA. The specificity of the method was tested on a collection of 61 reference plasmids and on 20 Salmonella enterica strains of different serotypes isolated in Italy. Results indicated that the inc/rep PCR method demonstrates high specificity and sensitivity in detecting replicons on reference plasmids and also revealed the presence of recurrent and common plasmids in epidemiologically unrelated Salmonella isolates of different serotypes. These results suggest that the method is potentially applicable to a large number of strains to trace the diffusion of specific multi-drug resistance plasmids in different environments.

AB - The epidemiological importance of tracing plasmids conferring drug resistance prompted us to develop a PCR method based on replicons (inc/rep PCR) of the major plasmid incompatibility groups among Enterobacteriaceae. Eighteen pairs of primers were designed to perform 5 multiplex- and 3 simplex-PCRs, recognizing FIA, FIB, FIC, HI1, HI2, I1-Iγ, L/M, N, P, W, T, A/C, K, B/O, X, Y, F, and FIIA. The specificity of the method was tested on a collection of 61 reference plasmids and on 20 Salmonella enterica strains of different serotypes isolated in Italy. Results indicated that the inc/rep PCR method demonstrates high specificity and sensitivity in detecting replicons on reference plasmids and also revealed the presence of recurrent and common plasmids in epidemiologically unrelated Salmonella isolates of different serotypes. These results suggest that the method is potentially applicable to a large number of strains to trace the diffusion of specific multi-drug resistance plasmids in different environments.

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ER -

Identification of plasmids by PCR-based replicon typing

Abstract

Bibliographical note

Keywords

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