title: Combined use of Pastorex Staph-Plus and either of two new chromogenic agars, MRSA ID and CHROMagar MRSA, for detection of methicillin-resistant Staphylococcus aureus.
abstract: We describe the search toward a fast and reliable strategy to detect and confirm the presence of methicillin-resistant Staphylococcus aureus (MRSA) in screening samples. First, we evaluated the sensitivities and specificities of oxacillin resistance screening agar (ORSA) with enrichment (tryptic soy broth [TSB] and ORSA [TSB-ORSA]) and without enrichment (ORSA), MRSA ID (MRSA_ID) plates, and CHROMagar MRSA (C_MRSA) plates, all of which were inoculated with equal volumes of a suspension made by emulsifying screening swabs. Whereas the sensitivities after 48 h were similar for all media tested (77% for MRSA_ID and ORSA; 73% for C_MRSA and ORSA after enrichment [TSB-ORSA]), the specificities of MRSA_ID (98% after 24 h and 94% after 48 h) and C_MRSA (98% after 24 h and 90% after 48 h) were superior to the specificities of ORSAs (92% after 24 h and 83% after 48 h) and TSB-ORSA (86% after 24 h and 81% after 48 h). Subsequently, the performance of the Pastorex Staph-Plus agglutination test with presumptive MRSA isolates taken directly from chromogenic agars (direct_Pastorex agglutination) was compared to that of the Pastorex Staph-Plus agglutination test with isolates from blood agar subcultures (conventional_Pastorex agglutination). When the direct_Pastorex agglutination test on MRSA_ID plates was combined with Gram staining, the direct_Pastorex agglutination test with samples from MRSA_ID plates was as reliable as the conventional_Pastorex agglutination test with samples from blood agar subcultures from MRSA_ID plates. In contrast, the direct_Pastorex agglutination test with samples from C_MRSA plates gave false-negative results. Finally, we calculated the processing times of the four different strategies, namely, (i) enrichment in TSB supplemented with NaCl, subsequent culture on ORSA, and the conventional_Pastorex agglutination test; (ii) direct inoculation of ORSA combined with conventional_Pastorex agglutination test; (iii) direct inoculation of MRSA_ID plates combined with Gram staining and the direct_Pastorex agglutination test; and (iv) direct inoculation of C_MRSA plates combined with Gram staining and the direct_Pastorex agglutination test. We concluded that the use of MRSA_ID in combination with Gram staining and the direct_Pastorex agglutination test is faster and more specific than the other strategies tested.
authors.affiliation: Department of Microbiology, Ghent University Hospital, De Pintelaan 185, B-9000 Ghent, Belgium.
authors.forename: Veerle
authors.initials: V
authors.lastname: Compernolle
authors.forename: Gerda
authors.initials: G
authors.lastname: Verschraegen
authors.forename: Geert
authors.initials: G
authors.lastname: Claeys
mesh_terms: D000362:Agar; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002863:Chromogenic Compounds / Q000378:metabolism*; D003470:Culture Media; D006801:Humans; D007841:Latex Fixation Tests; D008712:Methicillin / Q000494:pharmacology; D016106:Methicillin Resistance / Q000235:genetics; D008826:Microbial Sensitivity Tests / Q000379:methods; D016133:Polymerase Chain Reaction; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*
chemical_list: D000900:Anti-Bacterial Agents; D002863:Chromogenic Compounds; D003470:Culture Media; D000362:Agar; D008712:Methicillin