PCR-based identification of Staphylococcus epidermidis targeting gseA encoding the glutamic-acid-specific protease.
The frequency of the gseA gene encoding a glutamic acid-specific serine protease, GluSE, of Staphylococcus epidermidis was investigated. DNA hybridization analysis demonstrated that gseA existed exclusively in S. epidermidis but not in other bacteria examined. A single step PCR assay with a set of designed primers yielded amplification of gseA from all 69 clinical isolates of S. epidermidis taken from patients and healthy adults, whereas production of GluSE was observed in 74% (51/69) of the isolates. Furthermore, none of the 46 clinical isolates of other species of coagulase-negative staphylococci and 45 clinical isolates of Staphylococcus aureus showed amplification, except a Staphylococcus capitis strain. However, this strain was positive for a S. epidermidis-specific DNA region and the DNA sequence of the 16S rRNA gene showed 99% identity with that of S. epidermidis. Therefore, these results indicated that the present PCR assay for gseA was ubiquitous and highly specific for detection of S. epidermidis.
Ikeda
Y
Y
Department of Oral Microbiology, Iwate Medical University School of Dentistry, Morioka, Japan.
Ohara-Nemoto
Y
Y
Kimura
S
S
Ishibashi
K
K
Kikuchi
K
K
D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D001431:Bacteriological Techniques; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000032:analysis / Q000302:isolation & purification; D005798:Genes, Bacterial; D020459:Genes, rRNA; D006801:Humans; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D012697:Serine Endopeptidases / Q000032:analysis* / Q000235:genetics*; D013203:Staphylococcal Infections / Q000382:microbiology; D013210:Staphylococcus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D013211:Staphylococcus aureus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D013212:Staphylococcus epidermidis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase