Rapid Immunodiagnosis of Drug-Resistant Bacterial Pathogens

描述(申请人提供)：细菌对内酰胺类抗生素的耐药性正在威胁着限制这种最常用的抗生素的有效性。造成这一问题的一个主要因素是医生缺乏快速检测方法来识别 在开出特定的抗生素之前，对内酰胺类耐药的病原体进行检查。对于尿路感染(UTIs)，这是世界上最常见的细菌感染，尽管60%的泌尿病原体对这些药物敏感，但不使用较老的内酰胺类抗生素。对内酰胺类抗生素的耐药性是由内酰胺酶的表达产生的，内酰胺酶是一种使抗生素失活的酶。我们建议开发一种快速免疫检测试剂盒，用于检测各种内酰胺酶。在这个项目的第一阶段，我们的目标是开发针对最具临床问题的内酰胺酶-超广谱内酰胺酶(ESBLS)和碳青霉烯类酶的同源区域的交叉反应单抗(MAb)，然后开发一种能够区分内酰胺类耐药临床分离株和敏感分离株的夹心ELISA法。在第二阶段，我们会致力于 将这一实验室ELISA转化为一种坚固的、10分钟的侧向流动免疫检测试剂盒(类似于家庭怀孕测试)，能够直接在患者样本中检测内酰胺酶，从而检测内酰胺类耐药性。这一能力有望彻底改变目前的临床 尿路感染的治疗实践，允许甚至最早的内酰胺类药物的有效使用，并防止对当前一线治疗的耐药性选择。

DESCRIPTION (provided by applicant): Bacterial resistance to ¿-lactam antibiotics is threatening to limit the effectiveness of this most- used class of antibiotics. A major contributin factor to this problem is the lack of rapid testing methods for medical practitioners to identify ¿ lactam-resistant pathogens before prescribing specific antibiotics. For urinary tract infections (UTIs), the most common bacterial infections in the world, older ¿-lactam antibiotics are not used despite the fact that >60% of uropathogens are sensitive to these drugs. Resistance to ¿-lactam antibiotics is conferred by the expression of ¿-lactamases, enzymes that deactivate the antibiotics. We propose the development of a rapid immunoassay test kit for detection of any of a wide range of ¿-lactamases. In Phase I of this project, we aim to develop cross-reactive monoclonal antibodies (MAbs) targeting the homologous regions of the most clinically problematic classes of ¿-lactamases - extended- spectrum -lactamases (ESBLs) and carbapenemases - and then to develop a sandwich ELISA capable of distinguishing -lactam-resistant clinical bacterial isolates from ¿-lactam-sensitive isolates. In Phase II, we will aim to convert this laboratory ELISA into a robust, 10-minute lateral flow immunoassay test kit (similar to the home pregnancy test), capable of detecting ¿-lactamases, and therefore ¿-lactam resistance, directly in patient samples. This capability promises to revolutionize current clinical practice for treatment of (UTIs), allowing the effective use of even the earliest ¿-lactams and preventing the selection of resistance to current first-line treatments.