Dual Real-time PCR Detection of methicillin-resistant Staphylococcus aureus.docVIP

 PAGE \* MERGEFORMAT 24 Dual Real-time PCR Detection of methicillin-resistant Staphylococcus aureus 【Abstract】 Objective To establish the two heavy real-time polymerase chain reaction (duplex real  time PCR) Rapid detection of methicillin-resistant Staphylococcus aureus (MRSA). Methods The two re-SYBR Green real-time PCR for rapid detection of MRSA determining gene mecA and Staphylococcus aureus species-specific gene nuc, identified by melting curve analysis product. The results of the melting curve of all MRSA strains showed a mecA, nuc gene-specific peaks, methicillin-sensitive Staphylococcus aureus only nuc-feng, methicillin-resistant strains of the table Portuguese only mecA-feng, methicillin-sensitive tables Portuguese bacteria and other bacteria strains non-specific peaks; when the MRSA bacteria can be detected when the concentration of 102cfu/ml. In the 131 S. aureus in 30.53% (40/131) resistance; two heavy real-time PCR amplification of mecA gene 29.01% (38/131) positive, nuc gene 100% positive. Single primer and two heavy real-time PCR amplification of both the positive coincidence rate of 100%. Conclusion mecA, nuc genes of two re-real-time PCR can accurately and quickly identify methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci. Keywords: real-time polymerase chain reaction of methicillin-resistant Staphylococcus aureus mecA gene nuc gene Rapid detection of methicillin  resistant Staphylococcus aureus ABSTRACT Objective To establish a duplex real  time polymerase chain reaction (PCR) assay for rapid detection of methicillin  resistant Staphylococcus aureus (MRSA). Method A duplex SYBR Green real  time PCR assay targeting the mecA gene and a Staphylococcus aureus specific marker  nuc gene and identifying PCR products through melting curve analysis was used to rapidly identify MRSA. Results All MRSA strains tested in the study presented mecA and nuc gene specific peaks in the melting curv