Moussa Ihab, Shibl Atef M
Department of Pharmaceutics and Microbiology, College of Pharmacy, King Saud University, PO Box 2457, Riyadh 11451, Kingdom of Saudi Arabia.
Saudi Med J. 2009 May;30(5):611-7.
To examine the recovered strains phenotypically, by conventional methods and genotypically by polymerase chain reaction (PCR), for direct detection of Staphylococcus aureus (S. aureus) 16S ribosomal Ribonucleic Acid (rRNA) gene (which serves as an internal control) and mecA gene. Secondly, introduce multiplex PCR targeting at the same time S. aureus 16S rRNA, Panton-Valentine Leucocidin (PVL), and staphylococcal cassette chromosome mec (SCCmec) type IV.
Thirty-seven strains of S. aureus collected in 2007 from outpatient clinics in King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia, were tested in the College of Pharmacy phenotypically by conventional methods and genotypically by PCR for direct detection of S. aureus 16S rRNA and mecA genes. All the 37 strains, were tested also by multiplex PCR targeting at the same time S. aureus 16S rRNA, PVL, and (SCCmec) type IV.
Polymerase chain reaction detected all the 37 bacteriologically positive S. aureus (100%) and the mecA gene in all strains phenotypically resistant to methicillin (100%), at the same time it detected the mecA gene in 2 strains phenotypically sensitive to methicillin. Only 3 strains (8.1%) recovered from skin and soft tissue infections were positive for PVL and SCCmec type IV.
The PCR assay can be used for rapid detection of S. aureus and mecA gene. At the same time the multiplex PCR assay explained in this study is a rapid, sensitive, and reliable test for direct detection of community-acquired methicillin-resistant S. aureus.
通过传统方法对分离菌株进行表型检测,并通过聚合酶链反应(PCR)进行基因分型,以直接检测金黄色葡萄球菌(S. aureus)16S核糖体核糖核酸(rRNA)基因(用作内部对照)和mecA基因。其次,引入多重PCR,同时靶向金黄色葡萄球菌16S rRNA、杀白细胞素(PVL)和葡萄球菌盒式染色体mec(SCCmec)IV型。
2007年从沙特阿拉伯利雅得哈立德国王大学医院门诊收集的37株金黄色葡萄球菌,在药学院通过传统方法进行表型检测,并通过PCR进行基因分型,以直接检测金黄色葡萄球菌16S rRNA和mecA基因。所有37株菌株还通过多重PCR进行检测,同时靶向金黄色葡萄球菌16S rRNA、PVL和IV型(SCCmec)。
聚合酶链反应检测到所有37株细菌学阳性的金黄色葡萄球菌(100%)以及所有对甲氧西林表型耐药菌株中的mecA基因(100%),同时还在2株对甲氧西林表型敏感的菌株中检测到mecA基因。仅3株(8.1%)从皮肤和软组织感染中分离出的菌株PVL和SCCmec IV型呈阳性。
PCR检测可用于快速检测金黄色葡萄球菌和mecA基因。同时,本研究中解释的多重PCR检测是一种快速、灵敏且可靠的检测方法,用于直接检测社区获得性耐甲氧西林金黄色葡萄球菌。