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Journal of Clinical Microbiology, December 2000, p. 4351-4355, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Glyceraldehyde-3-Phosphate Dehydrogenase-Encoding Gene as a Useful Taxonomic Tool for Staphylococcus spp.

Javier Yugueros,1 Alejandro Temprano,1 Beatriz Berzal,1 María Sánchez,1 Carmen Hernanz,1 José María Luengo,2 and Germán Naharro1,*

Departamento de Sanidad Animal, Microbiología e Inmunología1 and Departamento de Bioquímica y Biología Molecular,2 Universidad de León, 24071 León, Spain

Received 22 June 2000/Returned for modification 4 September 2000/Accepted 24 September 2000

The gap gene of Staphylococcus aureus, encoding glyceraldehyde-3-phosphate dehydrogenase, was used as a target to amplify a 933-bp DNA fragment by PCR with a pair of primers 26 and 25 nucleotides in length. PCR products, detected by agarose gel electrophoresis, were also amplified from 12 Staphylococcus spp. analyzed previously. Hybridization with an internal 279-bp DNA fragment probe was positive in all PCR-positive samples. No PCR products were amplified when other gram-positive and gram-negative bacterial genera were analyzed using the same pair of primers. AluI digestion of PCR-generated products gave 12 different restriction fragment length polymorphism (RFLP) patterns, one for each species analyzed. However, we could detect two intraspecies RFLP patterns in Staphylococcus epidermidis, Staphylococcus hominis, and Staphylococcus simulans which were different from the other species. An identical RFLP pattern was observed for 112 S. aureus isolates from humans, cows, and sheep. The sensitivity of the PCR assays was very high, with a detection limit for S. aureus cells of 20 CFU when cells were suspended in saline. PCR amplification of the gap gene has the potential for rapid identification of at least 12 species belonging to the genus Staphylococcus, as it is highly specific.

* Corresponding author. Mailing address: Departamento de Sanidad Animal Microbiología e Immunología, Universidad de León, 24071 León, Spain. Phone: 34-87-291294. Fax: 34-87-291304. E-mail: dsagnc{at}unileon.es.

Journal of Clinical Microbiology, December 2000, p. 4351-4355, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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