| 实时荧光定量PCR检测耐甲氧西林金黄色葡萄球菌方法的建立与应用评价 |
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| 引用本文： | 牟晓峰,赵自云. 实时荧光定量PCR检测耐甲氧西林金黄色葡萄球菌方法的建立与应用评价[J]. 中华医院感染学杂志, 2011, 0(19): 4185-4187 |
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| 作者姓名： | 牟晓峰 赵自云 |
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| 作者单位： | 青岛市中心医院青岛大学医学院第二附属医院检验科,山东青岛,266042 |
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| 摘 要： | 目的利用实时荧光定量PCR技术,建立快速敏感特异的检测耐甲氧西林金黄色葡萄球菌(MRSA)的方法。方法以MRSA mecA基因为靶序列设计引物和探针,以自MRSA中提取的DNA为模板,优化引物、探针的浓度和退火温度,同时验证方法的特异性、敏感性、重复性、灵敏度和线性范围,并与K-B法和微量琼脂稀释法(MIC)相比较。结果建立的反应体系在上游引物、下游引物浓度为0.2 mmol/L、退火温度为55℃时,具有良好的特异性和敏感性,与大肠埃希菌等8种细菌均无交叉反应;对MRSA检测的灵敏度达到10 CFU/ml。结论建立的实时荧光PCR方法特异、灵敏、快速,能对MRSA做出快速准确的报告,实现准确、快速和定量检测。
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| 关 键 词： | 实时荧光定量PCR mecA基因 耐甲氧西林金黄色葡萄球菌 |
Establishment and clinical application of real-time PCR method for detection of methiciilin-resistant Staphylococcus aureus |
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| MU Xiao-feng,ZHAO Zi-yun. Establishment and clinical application of real-time PCR method for detection of methiciilin-resistant Staphylococcus aureus[J]. Chinese Journal of Nosocomiology, 2011, 0(19): 4185-4187 |
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| Authors: | MU Xiao-feng ZHAO Zi-yun |
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| Affiliation: | MU Xiao-feng,ZHAO Zi-yun(Qingdao Central Hospital,The Second Affiliated Hospital of Qingdao University Medical College,Qingdao,Shandong 266042,China) |
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| Abstract: | OBJECTIVE To establish a rapid,sensitive and specific real-time PCR method for detection of methicillin-resistant Staphylococcus aureus(MRSA).METHODS A pair of primers and a probe were designed depending on gene sequence of mecA of MRSA,and the DNA extracted from MRSA strains was used as a template to optimize primer and probe concentrations and annealing temperatures,and to verify the specificity,sensitivity,reproducibility and the linear range of the method,and compare with the K-B method and minimal inhi... |
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| Keywords: | Real-time quantitative PCR mecA MRSA |
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