Genomic DNA fingerprinting, using pulsed-field gel electrophoresis, of Staphylococcus intermedius isolated from dogs

Akira Shimizu From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan (Shimizu), and Department of Microbiology, Pathology, and Parasitology, College of Veterinary Medicine (Berkhoff), and Department of Genetics (Kloos, George, Ballard), North Carolina State University, Raleigh, NC 27695-7614.

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Herman A. Berkhoff From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan (Shimizu), and Department of Microbiology, Pathology, and Parasitology, College of Veterinary Medicine (Berkhoff), and Department of Genetics (Kloos, George, Ballard), North Carolina State University, Raleigh, NC 27695-7614.

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Wesley E. Kloos From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan (Shimizu), and Department of Microbiology, Pathology, and Parasitology, College of Veterinary Medicine (Berkhoff), and Department of Genetics (Kloos, George, Ballard), North Carolina State University, Raleigh, NC 27695-7614.

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Carol G. George From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan (Shimizu), and Department of Microbiology, Pathology, and Parasitology, College of Veterinary Medicine (Berkhoff), and Department of Genetics (Kloos, George, Ballard), North Carolina State University, Raleigh, NC 27695-7614.

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Deborah N. Ballard From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan (Shimizu), and Department of Microbiology, Pathology, and Parasitology, College of Veterinary Medicine (Berkhoff), and Department of Genetics (Kloos, George, Ballard), North Carolina State University, Raleigh, NC 27695-7614.

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Abstract

Objectives

To investigate the degree of polymorphism in the pulsed-field gel electrophoresis (PFGE) patterns of Staphylococcus intermedius and to assess the value of this typing method for discriminating strains.

Sample population

52 S intermedius isolates from diseased and healthy dogs.

Procedure

Chromosomal DNA of S intermedius was digested with restriction endonuclease Sma I, and the fragments were separated by PFGE in a 1 % agarose gel.

Results

Sma I cut the chromosomal DNA into 15 to 23 fragments ranging from about < 1 to 679 kb, and most of the detectable fragments were < 155 kb. Nine fragments, 115, 48, 33, 26, 16, 13, 10, 4, and < 1 kb, were shared by all or almost all (> 71 %) of the strains examined. Of the 52 strains, each had a different pattern. S intermedius had a high degree of restriction fragment length polymorphism. The PFGE patterns obtained for S intermedius were stable and reproducible when the strains were tested in the different experiments.

Conclusions

Genomic DNA fingerprinting by PFGE is an effective technique for discriminating S intermedius strains. The PFGE method appears to be a useful molecular marker for epidemiologic or ecologic studies of S intermedius. (Am J Vet Res 1996;57:1458-1462)

Abstract

Objectives

To investigate the degree of polymorphism in the pulsed-field gel electrophoresis (PFGE) patterns of Staphylococcus intermedius and to assess the value of this typing method for discriminating strains.

Sample population

52 S intermedius isolates from diseased and healthy dogs.

Procedure

Chromosomal DNA of S intermedius was digested with restriction endonuclease Sma I, and the fragments were separated by PFGE in a 1 % agarose gel.

Results

Sma I cut the chromosomal DNA into 15 to 23 fragments ranging from about < 1 to 679 kb, and most of the detectable fragments were < 155 kb. Nine fragments, 115, 48, 33, 26, 16, 13, 10, 4, and < 1 kb, were shared by all or almost all (> 71 %) of the strains examined. Of the 52 strains, each had a different pattern. S intermedius had a high degree of restriction fragment length polymorphism. The PFGE patterns obtained for S intermedius were stable and reproducible when the strains were tested in the different experiments.

Conclusions

Genomic DNA fingerprinting by PFGE is an effective technique for discriminating S intermedius strains. The PFGE method appears to be a useful molecular marker for epidemiologic or ecologic studies of S intermedius. (Am J Vet Res 1996;57:1458-1462)

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