Comparison of pulsed-field gel electrophoresis and phage typing for discriminating poultry strains of Staphylococcus aureus

Akira Shimizu From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan.

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Junichi Kawano From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan.

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Chikage Yamamoto From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan.

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Osamu Kakutani From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan.

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Manabu Fujita From the Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657, Japan.

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SUMMARY

Objective

To compare pulsed-field gel electrophoresis (PFGE) patterns of Staphylococcus aureus from chickens in England, Belgium, Bulgaria, Argentina, and Japan, to assess the value of PFGE for discriminating strains, and to compare results obtained by PFGE with those obtained by biotyping and phage typing.

Sample Population

78 S aureus isolates from diseased and healthy chickens.

Procedure

Chromosomal DNA of S aureus was digested with restriction endonuclease Sma l, and fragments were separated by PFGE in 1 % agarose gel.

Results

All 78 strains from 5 countries were classified as poultry ecovar according to a previously established biotyping system. Chromosomal DNA was cut by Sma l into 18 to 23 fragments ranging from about 3 to 685 kb. Seventy-eight strains produced 15 types, arbitrarily designated A to O, and 45 subtypes. Some differences were observed in PFGE patterns among countries. However, 10 fragments (333, 190, 110, 63, 55, 42, 34, 19, 10, and 3 kb) were highly conserved and were shared by almost all (> 78%) of the strains examined. The PFGE patterns were compared with those obtained by phage typing. All 29 strains belonging to avian phage-group II produced type A and 19 subtypes. Of the 15 strains belonging to phage-group I, 11 produced 8 types (B to H, O) and 5 subtypes that were different from those of type A.

Conclusions

Genomic DNA fingerprinting by PFGE is an effective technique for discriminating poultry S aureus strains and appears to be a useful method for subtyping strains of avian phage groups or the poultry-specific ecovar. (Am J Vet Res 1997;58:1412–1416)

SUMMARY

Objective

To compare pulsed-field gel electrophoresis (PFGE) patterns of Staphylococcus aureus from chickens in England, Belgium, Bulgaria, Argentina, and Japan, to assess the value of PFGE for discriminating strains, and to compare results obtained by PFGE with those obtained by biotyping and phage typing.

Sample Population

78 S aureus isolates from diseased and healthy chickens.

Procedure

Chromosomal DNA of S aureus was digested with restriction endonuclease Sma l, and fragments were separated by PFGE in 1 % agarose gel.

Results

All 78 strains from 5 countries were classified as poultry ecovar according to a previously established biotyping system. Chromosomal DNA was cut by Sma l into 18 to 23 fragments ranging from about 3 to 685 kb. Seventy-eight strains produced 15 types, arbitrarily designated A to O, and 45 subtypes. Some differences were observed in PFGE patterns among countries. However, 10 fragments (333, 190, 110, 63, 55, 42, 34, 19, 10, and 3 kb) were highly conserved and were shared by almost all (> 78%) of the strains examined. The PFGE patterns were compared with those obtained by phage typing. All 29 strains belonging to avian phage-group II produced type A and 19 subtypes. Of the 15 strains belonging to phage-group I, 11 produced 8 types (B to H, O) and 5 subtypes that were different from those of type A.

Conclusions

Genomic DNA fingerprinting by PFGE is an effective technique for discriminating poultry S aureus strains and appears to be a useful method for subtyping strains of avian phage groups or the poultry-specific ecovar. (Am J Vet Res 1997;58:1412–1416)

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