Identification of Staphylococcus aureus and determination of methicillin resistance directly from positive blood cultures by isothermal amplification and a disposable detection device.
A simple, rapid, and user-friendly procedure has been developed to identify Staphylococcus aureus and determine its methicillin resistance directly from gram-positive cocci in cluster-containing blood culture medium. The specimens were diluted and heated prior to amplification of the nuc and mecA genes with isothermal helicase-dependent amplification. Amplicons were detected using a disposable detection device. The analytical sensitivity of the assays was 50 CFU per reaction, and the clinical sensitivity and specificity were both 100% for S. aureus detection and 100% and 98% for methicillin resistance determination, respectively.
Goldmeyer
James
J
Molecular Infectious Disease Laboratory, Vanderbilt University Hospital, 4605 TVC, Nashville, TN 37232-5310, USA.
Li
Haijing
H
McCormac
Melinda
M
Cook
Steve
S
Stratton
Charles
C
Lemieux
Bertrand
B
Kong
Huimin
H
Tang
Wen
W
Tang
Yi-Wei
YW
D001426:Bacterial Proteins / Q000235:genetics; D001769:Blood / Q000382:microbiology; D003470:Culture Media; D004265:DNA Helicases / Q000378:metabolism; D004209:Disposable Equipment; D004720:Endonucleases / Q000235:genetics; D006801:Humans; D016106:Methicillin Resistance; D008836:Micrococcal Nuclease / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D046915:Penicillin-Binding Proteins; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000145:classification* / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification
D001426:Bacterial Proteins; D003470:Culture Media; D046915:Penicillin-Binding Proteins; D011933:Reagent Kits, Diagnostic; C487789:mecA protein, Staphylococcus aureus; C074002:nuc protein, staphylococcus; D004720:Endonucleases; D008836:Micrococcal Nuclease; D004265:DNA Helicases