Changing Cultures in Sepsis: Rapid single-cell pathogen identification and antibiotic susceptibility testing directly from whole blood

脓毒症中培养物的改变：直接从全血中进行快速单细胞病原体鉴定和抗生素敏感性测试

• 批准号: 10629223
• 负责人: Pak Kin Wong
• 金额: $ 73.03万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629223
• 关键词: Acceleration; Adopted; Antibiotic susceptibility; Antibiotics; Antimicrobial susceptibility; Bacteria; Bar Codes; Biological; Biological Assay; Blood Cells; Blood specimen; Cells; Characteristics; Classification; Clinical; Collaborations; Computer software; Cytology; Cytolysis; Data; Data Analyses; Detection; Diagnosis; Diagnostic; Disease; Drug Controls; Electroporation; Evolution; Goals; Growth; Hour; Individual; Industry; Kinetics; Label; Life; Measures; Membrane; Metabolism; Methods; Microbe; Microfluidic Microchips; Microfluidics; Microscopy; Minimum Inhibitory Concentration measurement; Modality; Molecular; Molecular Probes; Monitor; Morbidity - disease rate; Nanotubes; Pathogen detection; Patients; Performance; Phenotype; Predisposition; Process; Protocols documentation; RNA, Ribosomal, 18S; Research Personnel; Research Project Grants; Resistance; Resolution; Ribosomal RNA; Sampling; Scheme; Science; Sepsis; Sorting; Speed; System; Techniques; Technology; Temperature; Testing; Time; Translating; Triage; Validation; Vertebral column; Whole Blood; commensal microbes; commercialization; design; diagnostic platform; drug resistant pathogen; empowerment; fungus; image processing; imaging system; improved; innovation; microbial; microwave electromagnetic radiation; mortality; multidisciplinary; multimodality; novel; optical imaging; pathogen; precision medicine; product development; prospective; rapid diagnosis; response; single cell analysis; time use; validation studies

PROJECT SUMMARY Sepsis, commonly caused by bloodstream infections (BSI), is a rapidly progressive and life-threatening disease. Unfortunately, prolonged delay in microbiological diagnosis increases patient mortality, promotes the misuse of antibiotics, and consequently, the evolution of antibiotic-resistant pathogens. Herein, we aim to deliver an amplification-free, microfluidic system for pathogen detection, identification (ID), and antimicrobial susceptibility testing (AST) directly from whole blood. To achieve our goal, we propose a platform based on microfluidic- assisted microscopy to sort, trap, detect, and monitor pathogens at single cell resolution. For pathogen ID, we will adopt a multispectral barcoding scheme to differentially label molecular probes for direct multiplex ribosomal RNA (rRNA) detection to classify and speciate pathogens, along with a nanotube assisted microwave electroporation (NAME) technique to efficiently deliver the probes intracellularly for amplification-free single microbe detection. Positive pathogen ID will guide quantitative multimodal phenotypic AST (mPhAST), in which we will monitor early changes in microbial growth kinetics with cytological measures of viability in response to relevant antibiotic conditions at the single cell level to determine susceptibility/resistance with improved speed and reliability. Combined with upstream whole blood pre-processing for pathogen isolation and concentration followed by ID then AST, we aim to deliver sample to answer within 90 min for BSI triage and as early as 30 minutes more for antibiotic minimum inhibitory concentration (MIC) determination. We have assembled a superb team of multi-disciplinary investigators and industry-leading advisors with complementary expertise and a strong track record of collaboration. We propose the following aims: 1) to develop a rapid BSI triage protocol for broad pathogen detection, classification, and ID; 2) to develop a quantitative mPhAST; 3) to develop an integrated ID- mPhAST platform; 4) to perform analytical and clinical validation of our ID-mPhAST platform. Our short-term goal is to obtain the necessary preliminary data to plan for product development and commercialization, with the long-term goal of translating our diagnostic platform to reduce sepsis-related morbidity and mortality.

项目总结

项目成果

Association Between SARS-CoV-2 RNAemia and Post-Acute Sequelae of COVID-19.

SARS-CoV-2 RNAemia 与 COVID-19 急性后后遗症之间的关联。

• DOI: 10.1101/2021.09.03.21262934
• 发表时间: 2021
• 期刊: medRxiv : the preprint server for health sciences
• 作者: Ram-Mohan,Nikhil;Kim,David;Rogers,AngelaJ;Blish,CatherineA;Nadeau,KariC;Blomkalns,AndraL;Yang,Samuel
• 通讯作者: Yang,Samuel

A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections.

• DOI: 10.3390/bios11080288
• 发表时间: 2021-08-22
• 期刊: Biosensors
• 作者: Forsyth B;Torab P;Lee JH;Malcom T;Wang TH;Liao JC;Yang S;Kvam E;Puleo C;Wong PK
• 通讯作者: Wong PK

Using a 29-mRNA Host Response Classifier To Detect Bacterial Coinfections and Predict Outcomes in COVID-19 Patients Presenting to the Emergency Department.

• DOI: 10.1128/spectrum.02305-22
• 发表时间: 2022-12-21
• 期刊: Microbiology spectrum
• 影响因子: 3.7

Diagnosis of Bloodstream Infections: An Evolution of Technologies towards Accurate and Rapid Identification and Antibiotic Susceptibility Testing.

• DOI: 10.3390/antibiotics11040511
• 发表时间: 2022-04-12
• 期刊: Antibiotics (Basel, Switzerland)

Association Between SARS-CoV-2 RNAemia and Postacute Sequelae of COVID-19.

• DOI: 10.1093/ofid/ofab646
• 发表时间: 2022-03
• 期刊: Open forum infectious diseases
• 影响因子: 4.2
• 作者: Ram-Mohan N;Kim D;Rogers AJ;Blish CA;Nadeau KC;Blomkalns AL;Yang S
• 通讯作者: Yang S