Usefulness of Multiplex Real-Time PCR for Simultaneous Pathogen Detection and Resistance Profiling of Staphylococcal Bacteremia

• Published in: BioMed research international 2016-01, Vol.2016 (2016), p.1-7
• Main Authors: Kim, Hong Bin, Song, Junghan, Kim, Eui Chong, Kim, Eu Suk, Song, Kyoung-Ho, Hwang, Sang Mee, Park, Jeong Su, Hong, Yun Ji, Min, Young Gi, Kim, Taek Soo, Chung, Yousun, Park, Kyoung Un
• Format: Article
• Language: English
• Subjects: Bacteremia - diagnosis; Bacteremia - microbiology; Bacterial Load; Bacterial Typing Techniques - methods; Blood; Drug resistance in microorganisms; Drug Resistance, Bacterial - genetics; Genes; Health aspects; Humans; Laboratories; Medical examination; Methicillin-Resistant Staphylococcus aureus - classification; Methicillin-Resistant Staphylococcus aureus - genetics; Methicillin-Resistant Staphylococcus aureus - isolation & purification; Multiplex Polymerase Chain Reaction - methods; Nosocomial infections; Pathogens; Reproducibility of Results; RNA; Sensitivity and Specificity; Staphylococcal infections; Staphylococcal Infections - diagnosis; Staphylococcal Infections - microbiology; Staphylococcus aureus; Staphylococcus infections
• ISSN: 2314-6133; 2314-6141
• DOI: 10.1155/2016/6913860

Staphylococci are the leading cause of nosocomial blood stream infections. Fast and accurate identification of staphylococci and confirmation of their methicillin resistance are crucial for immediate treatment with effective antibiotics. A multiplex real-time PCR assay that targets mecA, femA specific for S. aureus, femA specific for S. epidermidis, 16S rRNA for universal bacteria, and 16S rRNA specific for staphylococci was developed and evaluated with 290 clinical blood culture samples containing Gram-positive cocci in clusters (GPCC). For the 262 blood cultures identified to the species level with the MicroScan WalkAway system (Siemens Healthcare Diagnostics, USA), the direct real-time PCR assay of positive blood cultures showed very good agreement for the categorization of staphylococci into methicillin-resistant S. aureus (MRSA), methicillin-susceptible S. aureus (MSSA), methicillin-resistant S. epidermidis (MRSE), methicillin-susceptible S. epidermidis (MSSE), methicillin-resistant non-S. epidermidis CoNS (MRCoNS), and methicillin-susceptible non-S. epidermidis CoNS (MSCoNS) ( κ = 0.9313 ). The direct multiplex real-time PCR assay of positive blood cultures containing GPCC can provide essential information at the critical point of infection with a turnaround time of no more than 4 h. Further studies should evaluate the clinical outcome of using this rapid real-time PCR assay in glycopeptide antibiotic therapy in clinical settings.