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Chapter 50 : Molecular Typing of by Pulsed-Field Gel Electrophoresis and Amplified Fragment Length Polymorphism Analysis

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Molecular Typing of by Pulsed-Field Gel Electrophoresis and Amplified Fragment Length Polymorphism Analysis, Page 1 of 2

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Abstract:

When detailed epidemiologic characterization is required, standardized molecular typing methods are indispensable for the subtyping of serogroups. Therefore, a comparison was made between pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP) analysis for genotyping of , using standardized protocols from the European Working Group on Legionella Infections (EWGLI). DNA fingerprint patterns derived from AFLP seem to be superior to the patterns derived from PFGE, in both speed and interpretability. AFLP and PFGE have similar discriminatory power. AFLP and PFGE are powerful tools for the epidemiological subtyping of . The authors emphasize the importance of the standardization of data analysis and data exchange using the latest software (BioNumerics). Studies are under way to determine both the intralaboratory and interlaboratory reproducibility more extensively for further standardization of these powerful subtyping methods used by an increasing number of European countries.

Citation: Wannet W, van der Zwaluw W, Heck M, Elzenaar C, Maas H, Brunings H, Schellekens J, Bergmans A, van der Zee A, Thijssen E, Peeters M. 2002. Molecular Typing of by Pulsed-Field Gel Electrophoresis and Amplified Fragment Length Polymorphism Analysis, p 260-262. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch50

Key Concept Ranking

Pulsed-Field Gel Electrophoresis
1.1829933
Legionella pneumophila
0.8333333
1.1829933
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Figures

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FIGURE 1

Case example showing the application of PFGE and AFLP for the molecular subtyping of

Citation: Wannet W, van der Zwaluw W, Heck M, Elzenaar C, Maas H, Brunings H, Schellekens J, Bergmans A, van der Zee A, Thijssen E, Peeters M. 2002. Molecular Typing of by Pulsed-Field Gel Electrophoresis and Amplified Fragment Length Polymorphism Analysis, p 260-262. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch50
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References

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1. Fry, N. K.,, S. Alexiou-Daniel,, J. M. Bangsborg,, S. Bernander,, M. Castellani Pastoris,, J. Etienne,, B. Forsblom,, V. Gaia,, J. H. Helbig,, D. Lindsay,, P. C. Luck,, C. Pelaz,, S. Uldum,, and T. G. Harrison. 1999. A multicenter evaluation of genotypic methods for the epidemiologic typing of Legionella pneumophila serogroup 1: results of a pan-European study. Clin. Miaobiol. Infect. 5: 462477.
2. Fry, N. K.,, J. M. Bangsborg,, S. Bernander,, J. Etienne,, B. Forsblom,, V. Gaia,, P. Hasenberger,, D. Lindsay,, A. Papoutsi,, C. Pelaz,, M. Struelens,, S. A. Uldum,, P. Visca,, and T. G. Harrison. 2000. Assessment of intercentre reproducibility and epidemiological concordance of Legionella pneumophila serogroup 1 genotyping by amplified fragment length polymorphism analysis. Eur. J. Clin. Miaobiol. Infect. Dis. 19:773780.
3. Riffard, S.,, F. Lo Presti,, F. Vendenesch,, F. Forey,, M. Reyrolle,, and J. Etienne. 1998. Comparative analysis of infrequent-restriction-site PCR and pulsed-field gel electrophoresis for epidemiological typing of Legionella pneumophila serogroup 1 strains. J. Clin. Microbiol. 36:161167.
4. Valsangiacomo, C.,, F. Baggi,, V. Gaia,, T. Balmelli,, R. Peduzzi,, and J. C. Piffaretti. 1995. Use of amplified fragment length polymorphism in molecular typing of Legionella pneumophila and application to epidemiological studies. J. Clin. Miaobiol. 33:17161719.

Tables

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TABLE 1

Subdivision of PFGE types in AFLP types and vice versa

Citation: Wannet W, van der Zwaluw W, Heck M, Elzenaar C, Maas H, Brunings H, Schellekens J, Bergmans A, van der Zee A, Thijssen E, Peeters M. 2002. Molecular Typing of by Pulsed-Field Gel Electrophoresis and Amplified Fragment Length Polymorphism Analysis, p 260-262. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch50

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