title: Direct mecA detection from blood culture bottles by branched-DNA signal amplification.
abstract: A branched-DNA (bDNA) signal amplification method was used to detect the mecA gene directly from blood culture broth growing staphylococci. BACTEC blood culture bottles with positive growth indices and containing staphylococcus-like organisms as shown by Gram stain were tested for the presence of the mecA gene. Comparison of test results was done among 225 patients (one blood culture from each patient). Compared with PCR, the sensitivity and specificity of the bDNA method are 100 and 99%, respectively. The bDNA test is carried out in a 96-well format and requires approximately 6 h to perform. Our preliminary results suggest that direct detection of the mecA gene by bDNA signal amplification is (i) sensitive enough to detect mecA directly from blood culture bottles without the requirement for subculture and (ii) as sensitive and specific as the PCR-based method.
authors.affiliation: Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic and Foundation, Rochester, Minnesota.
authors.forename: X
authors.initials: X
authors.lastname: Zheng
authors.forename: C P
authors.initials: CP
authors.lastname: Kolbert
authors.forename: P
authors.initials: P
authors.lastname: Varga-Delmore
authors.forename: J
authors.initials: J
authors.lastname: Arruda
authors.forename: M
authors.initials: M
authors.lastname: Lewis
authors.forename: J
authors.initials: J
authors.lastname: Kolberg
authors.forename: F R
authors.initials: FR
authors.lastname: Cockerill
authors.forename: D H
authors.initials: DH
authors.lastname: Persing
mesh_terms: D016470:Bacteremia / Q000175:diagnosis / Q000382:microbiology; D001426:Bacterial Proteins / Q000235:genetics*; D001769:Blood / Q000382:microbiology*; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000175:diagnosis* / Q000382:microbiology; D013210:Staphylococcus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D004269:DNA, Bacterial; C083616:mecA protein, Bacillus subtilis