Point-of-care antimicrobial susceptibility testing based on simultaneous tracking of multi-phenotypic features of single bacterial cells

摘要 抗生素耐药性已成为一个重大的公共卫生威胁。为了解决这个问题，一种快速的病原体 需要鉴定（ID）和抗菌药物敏感性测试（AST）技术来提供及时的诊断， 在初级卫生保健环境中监测耐药性感染和提供准确的抗生素治疗，包括- 医院和护理点（POC）。本项目旨在开发一种即时护理AST（POCASTTM） 基于大图像体积显微镜技术的技术，能够直接检测个体 在没有培养或病原体分离的情况下在临床样品中检测细菌细胞，以及机器学习模型， 可以快速确定病原体和易感性。为了建立该技术，该项目将侧重于 尿路感染（UTIs）。UTI每年影响数百万人，而通常引起UTI的病原体 UTI是对包括碳青霉烯类在内的抗生素耐药性构成最大威胁的微生物， 耐药肠杆菌科（CRE）和产超广谱β-内酰胺酶（ESBL）的肠杆菌， 科。 该项目将重点关注：1）开发大图像体积显微镜和机器学习模型， 直接在患者尿样中同时追踪单个细菌细胞的多表型特征，以及 对UTI进行快速自动病原体ID和AST; 2）建立原型仪器; 3）验证 使用大规模临床样本的UTI仪器。技术的成功开发和验证- nology将在患者就诊当天提供精确的抗生素处方。 该项目将由一个具有生物传感器专业知识的多学科团队（生物设计中心）进行。 生物电子学和生物传感器，亚利桑那州立大学），微生物学和传染病（生物设计中心免疫， 治疗，疫苗和病毒治疗，ASU），生物医学仪器开发和生产（生物传感 Instrument Inc.），和临床试验（临床微生物学实验室，马约诊所）。 !

ABSTRACT Antibiotic resistance has become a significant public health threat. To combat the problem, a rapid pathogen identification (ID) and antimicrobial susceptibility testing (AST) technology is needed to provide timely diagno- sis of resistant infections and delivery of accurate antibiotic treatment at primary health-care settings, includ- ing hospitals and point-of-care (POC). The present project aims to develop a point-of-care AST (POCASTTM) technology based on a large-image-volume microscopy technique that enables direct detection of individual bacterial cells in clinical samples without culturing or pathogen isolation, and a machine-learning model that allows fast determination of pathogen and susceptibility. To establish the technology, the project will focus on urinary tract infections (UTIs). UTIs affect millions of people annually, and the pathogens that usually cause UTIs are the organisms that pose the highest threat of antimicrobial resistance, including carbapenem- resistant Enterobacteriaceae (CRE) and extended spectrum β-lactamase (ESBL)-producing Enterobacteri- aceae. This project will focus on: 1) developing the large-image-volume microscopy and machine learning model for simultaneous tracking of multi-phenotypic features of single bacterial cells directly in patient urine sample, and performing rapid automatic pathogen ID and AST for UTIs; 2) building prototype instrument, and 3) validating the instrument for UTIs using large scale clinical samples. Successful development and validation of the tech- nology will enable precise antibiotic prescription on the same day of patient visit. The project will be carried out by a multidisciplinary team with expertise in biosensors (Biodesign Center for Bioelectronics and Biosensors, ASU), microbiology and infectious diseases (Biodesign Center for Immuno- therapy, Vaccines and Virotherapy, ASU), biomedical instrument development and production (Biosensing Instrument Inc.), and clinical testing (Clinical Microbiology Laboratory, Mayo Clinic). !