GENETIC AND METABOLIC PROBES FOR ANTIBIOTIC RESISTANCE

目的是开发一套快速、可靠、半自动化的DNA探针 耐药葡萄球菌属的检测方法 各种院内感染。 首先，我们将开发一种DNA探针测试， 对耐甲氧西林链球菌的mecA基因进行了序列分析。金黄色葡萄球菌（MRSA）和凝固酶- 阴性葡萄球菌属（MRCNS）。 mecA基因编码 MRSA和MRCNS分离株的甲氧西林耐药率为97%-100%。 直接 mecA的检测将比目前更快速、准确和可靠 培养依赖性甲氧西林敏感性试验， 通过体外抗性表型的高度可变表达。 的 DNA探针测试还将包括用于识别个体的探针 葡萄球菌属，允许同时进行菌种鉴定和检测 甲氧西林耐药性的测试 测试将于 从初级分离平板中挑选菌落，仅需要15个 35分钟的人工处理，然后是35分钟的自动处理 处理，在1小时内产生结果。 我们还将开发检测 用于治疗MRSA和MRCNS的二线抗生素的耐药性 感染. 这些检测将在确认为 通过mecA/菌种联合试验确定甲氧西林耐药，并将需要 2-4小时，以获得结果，在30分钟的手动处理。 整套检测将产生完整的菌种ID和抗生素 在从平板上挑取菌落的半天内得出敏感性结果。

The goal is to develop a suite of rapid, reliable, semiautomated DNA probe assays for antibiotic-resistant Staphylococcus species responsible for a variety of nosocomial infections. First, we will develop a DNA probe test for the mecA gene of methicillin-resistant S. aureus (MRSA) and coagulase- negative Staphylococcus species (MRCNS). The mecA gene codes for methicillin resistance in 97% to 100% of MRSA and MRCNS isolates. Direct detection of mecA will be more rapid, accurate, and reliable than current culture-dependent methicillin susceptibility tests, which are compromised by highly variable expression of the resistance phenotype in vitro. The DNA probe test will also include probes for identification of individual Staphylococcus species, allowing simultaneous species ID and detection of methicillin resistance in a single test. The test will be conducted on colonies picked from primary isolation plates, and will require only 15 minutes of manual processing followed by 35 minutes of automated processing, yielding results in under 1 hour. We will also develop assays for resistance to second-line antibiotics used to treat MRSA and MRCNS infections. These tests will be conducted on isolates confirmed to be methicillin resistant by the combined mecA/species assay, and will require 2-4 hours to obtain results with under 30 minutes of manual processing. The entire suite of assays will yield complete species ID and antibiotic susceptibility results within half a day of picking colonies from plates.