Cloney L, Marlowe C, Wong A, Chow R, Bryan R
ID Biomedical Corporation, 8855 Northbrook Court, Burnaby BC, V5J 5J1, Canada.
Mol Cell Probes. 1999 Jun;13(3):191-7. doi: 10.1006/mcpr.1999.0235.
A novel method has been developed for the detection of the mecA gene, that confers the principle mechanism of methicillin resistance in staphylococci. Cycling Probe Technology (CPT) is a rapid, simple, isothermal method for the detection of specific target sequences. CPT utilizes a unique chimeric DNA-RNA-DNA probe sequence that provides an RNase H sensitive scissile link when hybridized to a complementary target DNA sequence. In the presence of target DNA, the cycling reaction converts full-length chimeric probe into cleaved probe fragments, which accumulate and are quantified. A cycling probe designed for detection of a specific sequence within the mecA gene was used to develop a culture confirmation assay for methicillin resistant Staphylococcus aureus. The CPT assay was used to screen 238 S. aureus isolates and the results were in complete agreement with detection of the mecA gene by polymerase chain reaction (PCR). Detection of mecA should be considered the gold standard for determining methicillin resistance in S. aureus. This study demonstrates the feasibility of using CPT to meet this need.
已开发出一种用于检测mecA基因的新方法，该基因赋予葡萄球菌对甲氧西林耐药的主要机制。循环探针技术（CPT）是一种用于检测特定靶序列的快速、简单的等温方法。CPT利用独特的嵌合DNA-RNA-DNA探针序列，当与互补靶DNA序列杂交时，该序列提供对核糖核酸酶H敏感的可切割连接。在存在靶DNA的情况下，循环反应将全长嵌合探针转化为切割后的探针片段，这些片段会积累并进行定量。为检测mecA基因内特定序列而设计的循环探针被用于开发耐甲氧西林金黄色葡萄球菌的培养确认试验。CPT试验用于筛选238株金黄色葡萄球菌分离株，结果与通过聚合酶链反应（PCR）检测mecA基因完全一致。检测mecA应被视为确定金黄色葡萄球菌对甲氧西林耐药性的金标准。本研究证明了使用CPT满足这一需求的可行性。
