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Chapter 46 : Molecular Typing of Strains with Pulsed-Field Gel Electrophoresis and Random Primer-Amplified Polymorphic DNA in Nosocomial Legionnaires' Disease

Authors: Laura Franzin1, Daniela Cabodi1, Alessandro Sinicco1, Giovanni Di Petri1
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Affiliations: 1: Infectious Diseases Unit, University of Turin, Turin 10149, Italy
Content Type: Monograph
Publication Year: 2002

Category: Bacterial Pathogenesis

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Molecular Typing of Strains with Pulsed-Field Gel Electrophoresis and Random Primer-Amplified Polymorphic DNA in Nosocomial Legionnaires' Disease, Page 1 of 2

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Abstract:

This chapter investigates the epidemiological relatedness between strains, isolated from a patient with Legionnaires' disease and from the hospital water supply. Strains of serogroup 1 isolated from the patient and from the hospital environment, and clinical and environmental unrelated strains were typed by pulsed-field gel electrophoresis (PFGE) and by random amplified polymorphic DNA PCR (RAPD-PCR). serogroup 1 strains isolated from the patient and 11 strains isolated from hot water samples collected from different sites of the hospital water supply showed the same profile by PFGE with l and l. These strains shared identical profiles also by RAPD-PCR with the two primers used. The profiles of serogroup 1 Philadelphia 1 type strain and of the clinical and environmental unrelated strains were different from the above strains by PFGE and by RAPD-PCR. The molecular typing results demonstrated that the strains of serogroup 1, isolated from the patient and from the hospital hot water, were indistinguishable and thus genetically related, showing that the hospital water was the source of infection and thus confirming the nosocomial origin.

Citation: Franzin L, Cabodi D, Sinicco A, Di Petri G. 2002. Molecular Typing of Strains with Pulsed-Field Gel Electrophoresis and Random Primer-Amplified Polymorphic DNA in Nosocomial Legionnaires' Disease, p 248-250. 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.ch46
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Molecular Typing of Legionella Strains with Pulsed-Field Gel Electrophoresis and Random Primer-Amplified Polymorphic DNA in Nosocomial Legionnaires' Disease
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Citation: Franzin L, Cabodi D, Sinicco A, Di Petri G. 2002. Molecular Typing of Strains with Pulsed-Field Gel Electrophoresis and Random Primer-Amplified Polymorphic DNA in Nosocomial Legionnaires' Disease, p 248-250. 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.ch46
/content/10.1128/9781555817985.chap46.fig46-1
FIGURE 1

PFGE of I-cleaved genomic DNA of serogroup 1 strains. Lanes: 1, patient's isolate; 2 through 8, environmental isolates from different sites of the hospital water supply; 11, serogroup 1 Philadelphia 1; 12 and 13, clinical unrelated isolates; 14 and 15, environmental unrelated isolates. Lambda DNA concatamers (48.5 kb) (L, lane 10) and yeast chromosomal DNA of (S, lane 9) were used as molecular size standards.

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10.1128/9781555817985/fig46-1.gif
Image of FIGURE 1
FIGURE 1

PFGE of I-cleaved genomic DNA of serogroup 1 strains. Lanes: 1, patient's isolate; 2 through 8, environmental isolates from different sites of the hospital water supply; 11, serogroup 1 Philadelphia 1; 12 and 13, clinical unrelated isolates; 14 and 15, environmental unrelated isolates. Lambda DNA concatamers (48.5 kb) (L, lane 10) and yeast chromosomal DNA of (S, lane 9) were used as molecular size standards.

Citation: Franzin L, Cabodi D, Sinicco A, Di Petri G. 2002. Molecular Typing of Strains with Pulsed-Field Gel Electrophoresis and Random Primer-Amplified Polymorphic DNA in Nosocomial Legionnaires' Disease, p 248-250. 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.ch46
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References

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6. Strulens, M. J.,, N. Maes,, F. Rost,, A. Deplano,, F. Jacobs,, C. Liesnard,, N. Bornstein,, F. Grimont,, S. Lauwers,, M. P. McIntyre,, and E. Serruys. 1992. Genotypic and phenotypic methods for the investigation of a nosocomial Legionella pneumophila outbreak and efficacy of control measures. J. Infect. Dis. 166: 22– 30.
7. Tenover, F. C.,, R. D. Arbeit,, R. V. Goering,, P. A. Mickelsen,, B. E. Murray,, D. H. Persing,, and B. Swaminathan. 1995. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J. Clin. Microbiol. 33: 2233– 2239.
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9. Wilkinson, H. W. 1987. Hospital-Laboratory Diagnosis of Legionella Infections. U.S. Department of Health and Human Services. Centers for Disease Control,Atlanta,Ga..

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