Rapid and sensitive detection of the femA gene in staphylococci by enzymatic detection of polymerase chain reaction (ED-PCR): comparison with standard PCR analysis.
Despite its recent cloning and characterization, the physiological role of FemA remains unclear. To easily and reliably identify femA in staphylococci, we analysed 45 blood isolates of staphylococci using enzymatic detection of polymerase chain reaction (ED-PCR). ED-PCR is a new method that detects amplified PCR products using biotin-streptavidin affinity and an enzyme-linked antibody. Of 45 samples, 34 strains contained the femA gene as detected by ED-PCR. Phenotyping analysis showed that these 34 strains (femA-positive) were Staphylococcus aureus and that the other 11 (femA-negative) were S. epidermidis. These results were completely consistent with the results of femA detection using standard PCR and subsequent Southern blot hybridization. The ED-PCR procedure was complete within 4 h and could be done in one tube. We conclude that ED-PCR is a rapid, simple and reliable method for detecting the femA gene and that it provides an important means of classifying staphylococcal strains in the clinical field.
Kizaki
M
M
Division of Hematology, School of Medicine, Keio University, Tokyo, Japan.
Kobayashi
Y
Y
Ikeda
Y
Y
D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers; D005798:Genes, Bacterial / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D013211:Staphylococcus aureus / Q000145:classification / Q000235:genetics*; D013212:Staphylococcus epidermidis / Q000145:classification / Q000235:genetics*
D017931:DNA Primers