Rapid detection of methicillin-resistant staphylococci from blood culture bottles by using a multiplex PCR assay.
Rapid detection and accurate identification of methicillin-resistant staphylococci are critical for the effective management of infections caused by these organisms. We describe a multiplex PCR-based assay for the direct detection of methicillin-resistant staphylococci from blood culture bottles (BacT/Alert; Organon-Teknika, Durham, N.C.). A simple lysis method followed by a multiplex PCR assay designed to detect the nuc, mecA, and bacterial 16S rRNA genes was performed. A total of 306 blood culture specimens were collected over a period of 10 months from June 1998 to April 1999, consisting of 236 blood cultures growing staphylococci (including 124 methicillin-resistant Staphylococcus spp.), 50 positive blood cultures which grew organisms other than staphylococci, and 20 blood cultures that were negative for bacterial and fungal pathogens after 5 days of incubation and terminal subculture. DNA extraction, PCR, and detection could be completed in 2.5 h. Of the positive blood cultures with staphylococci, the multiplex PCR assay had a sensitivity and specificity of 99.2% and 100%, respectively. Our results show that rapid, direct detection of methicillin-resistant staphylococci is possible, allowing clinicians to make prompt and effective decisions for the management of patients with staphylococcal bacteremia.
Louie
L
L
Sunnybrook and Women's College Health Sciences Centre, Toronto, Ontario, Canada. lisa.louie@swchsc.on.ca
Goodfellow
J
J
Mathieu
P
P
Glatt
A
A
Louie
M
M
Simor
A E
AE
D016470:Bacteremia / Q000382:microbiology*; D001426:Bacterial Proteins; D001769:Blood / Q000382:microbiology*; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis; D004720:Endonucleases / Q000235:genetics; D006801:Humans; D016106:Methicillin Resistance; D008836:Micrococcal Nuclease; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000382:microbiology; D013210:Staphylococcus / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D013997:Time Factors
D001426:Bacterial Proteins; D003470:Culture Media; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; C074002:nuc protein, staphylococcus; D004720:Endonucleases; D008836:Micrococcal Nuclease