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首页> 外文会议>12th Symposium on Bioluminescence and Chemiluminescence, Apr 5-9, 2002, Robinson College, University of Cambridge, UK >RAPID DETECTION OF MRSA FROM CLINICAL SAMPLES USING MAGNETIC SEPARATION AND AK BIOLUMINESCENCE
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RAPID DETECTION OF MRSA FROM CLINICAL SAMPLES USING MAGNETIC SEPARATION AND AK BIOLUMINESCENCE

机译：磁性分离和AK生物发光技术从临床样品中快速检测MRSA

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摘要

Staphylococcus aureus is an opportunistic pathogen that occurs as part of the normal body flora in about a quarter of healthy adults. However, its methicillin-resistant variant (MRSA) has become endemic in hospitals worldwide, causing wound sepsis and other infections, particularly post-operatively. Patients are tested for MRSA by taking swabs, usually from the nose, perineum, armpit or wound. Analysed using conventional microbiological techniques, presumptive negative results are obtained in two days and positive results in two to four days. A prototype test for rapid MRSA detection was developed that used a combination of magnetic separation and adenylate kinase bioluminescence. The technique employed a short culture step (3-4 h) in a methicillin-containing broth to lyse susceptible bacteria prior to capturing resistant Staph. aureus cells using fibrinogen-coated magnetic particles. Staph. aureus has an affinity for fibrinogen that is not shown by other closely related species. Patient swabs were used to inoculate small volumes of broth containing methicillin, which were then incubated for 3 h. Previous experiments had shown that effective cell lysis of susceptible bacterial species could be obtained within this time. The fibrinogen-coated paramagnetic beads were then used to capture intact (i.e. antibiotic-resistant) cells, washed to remove extracellular adenylate kinase (AK) and then assayed using AK bioluminescence. The total assay time was approximately 4 h.
机译：金黄色葡萄球菌是一种机会病原体，在大约四分之一的健康成年人中作为正常人体菌群的一部分出现。然而，其耐甲氧西林的变异体（MRSA）已在世界各地的医院中流行，引起伤口败血症和其他感染，尤其是在术后。通常通过从鼻子，会阴部，腋窝或伤口拭子对患者进行MRSA检测。使用常规微生物技术进行分析，在两天内获得假定的阴性结果，在两到四天内获得阳性结果。开发了用于快速MRSA检测的原型测试，该测试结合了磁分离和腺苷酸激酶生物发光技术。该技术在捕获抗性葡萄球菌之前，先在含有甲氧西林的肉汤中进行短暂的培养步骤（3-4小时），以裂解易感细菌。金黄色葡萄球细胞使用纤维蛋白原包被的磁性颗粒。葡萄球菌。金黄色葡萄球菌对纤维蛋白原具有亲和力，而其他紧密相关的物种却没有。患者拭子用于接种少量含甲氧西林的肉汤，然后孵育3小时。先前的实验表明，可以在这段时间内对敏感细菌进行有效的细胞裂解。然后将纤维蛋白原包被的顺磁珠用于捕获完整的（即抗生素抗性）细胞，洗涤以除去细胞外腺苷酸激酶（AK），然后使用AK生物发光法进行分析。总测定时间约为4小时。

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