Comparison of genotypic and phenotypic methods for species identification of coagulase-negative staphylococcal isolates from blood cultures.
Coagulase-negative staphylococci are often isolated from blood cultures. Simple methods are needed for correct identification of those species most frequently found. In this study, PCR methods were developed for the identification of S. epidermidis and S. haemolyticus, based on DNA sequences in their 16S rDNA. The results obtained by these methods were compared with those obtained using a number of phenotypic methods, including two commercial kits API Staph and Staphzyme. Fifteen type collection strains and 133 blood culture isolates were tested. The sensitivity of the PCR for identification of S. epidermidis was 99% compared with API Staph, but the specificity was lower, 94%, because of positive results also for S. capitis. The results by the PCR for identification of S. haemolyticus correlated closely with the Staphzyme results, 13 isolates being identified by Staphzyme and 16 by the PCR. API Staph, however, identified only four clinical isolates as S. haemolyticus, probably too few. Among the individual phenotypic tests performed, a trehalose-mannitol agar method and a desferrioxamine disc diffusion test for the identification of S. epidermidis were found to be very accurate. Anaerobic growth after overnight incubation could be used to distinguish S. epidermidis from S. hominis. The conclusion is that a majority of all Gram-positive, catalase-positive and coagulase-negative blood culture isolates can be typed as regards species level using only a few genotypic and/or phenotypic tests.
Hedin
G
G
Department of Clinical Microbiology, Uppsala University Hospital, Sweden.
D015373:Bacterial Typing Techniques; D001483:Base Sequence; D001769:Blood; D003030:Coagulase / Q000378:metabolism; D004269:DNA, Bacterial; D005838:Genotype; D008837:Micrococcus / Q000145:classification; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D013210:Staphylococcus / Q000145:classification* / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification
D003030:Coagulase; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D011933:Reagent Kits, Diagnostic