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Journal of Clinical Microbiology, December 2000, p. 4351-4355, Vol. 38, No. 12
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.
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.
*
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.
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