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Culture-Independent Pathogen Identification Through Rapid Lipidomic Profiling

Award Number: R01AI198756
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 09/08/2026
PERIOD OF PERFORMANCE END DATE: 08/31/2031

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Culture-Independent Pathogen Identification Through Rapid Lipidomic Profiling - PROJECT SUMMARY Rapid diagnostic tests (RDTs) have become essential in clinical microbiology laboratories due to their ability to optimize the management of urinary tract, bloodstream, and cerebrospinal fluid infections, leading to better patient outcomes including reduced mortality and morbidity. Among these, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) stands out for its rapid, accurate, and cost-effective identification of pathogens, utilizing a comprehensive mass spectral library from ribosomal proteins, highly conserved in bacterial species for proteomic analysis.The major drawbacks of these protein-based platforms are their limited capabilities in detecting polymicrobial infections and antimicrobial resistance, and their inability to conduct direct sample analysis without the required time (generally 24 hours) to produce a pure colony by culture. To overcome these issues, we have developed a novel lipid extraction method, Fast Lipid Analysis Technique (FLAT), which mimics identification from a single colony like proteomic methods but provides identifications directly from a specimen without ex vivo growth. FLAT extracts lipids directly on a MALDI plate in less than 1 h, significantly reducing labor and handling time compared to those of previous lipid analysis methods while providing identification of microbes and antimicrobial resistance directly from complex biological samples. For this application we will develop: 1) a multimodal platform (FLAT) for rapid microbe identification from in vitro or in vivo mono- or poly-microbial samples; 2) an automated lipid and O-antigen structure annotation system using reference Kendrick Mass Defect on TOF mass analyzers together with tandem MS in days versus weeks; 3) novel technologies (Panini Press and SuperFLAT plates) that improve rigor by improving reliability and limit of detection (LOD) from a single colony (~105-6 cells) to 103 microbes, respectively; 4) a rapid serovar assignment method called FLOAT (Fast Lipid/O-Antigen Test), which is a derivation of the FLAT method, that detects O- antigens from Gram-negative organisms along with their released lipid A and any other lipids if the organism is not Gram-negative; 5) PostalService, a machine learning platform automating microbial identification and serovar assignment via high-throughput MALDI-TOF MS data; and 6) extending the FLAT platform through the use of internal standards to allow CFU-like values to be reported for direct analysis of urinary tract infections, blood, and CSF samples using the FDA-approved Bruker Biotyper sirius platform without culture. Together, these advances aim to revolutionize microbial lipidomics and diagnostics through enhanced detection, standardized quantification, and machine learning-driven pattern recognition. This application will ultimately produce a lipid-based identification approach that can accurately and rapidly identify pathogens from real-world biological samples without culture for research use. Importantly, FLAT is easily adoptable by other laboratories where we expect innovative and rapid advances in understanding the role of microbial membrane lipids in a wide variety of clinical and research applications.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2026 ( Subtotal = $769,725 )
2026	2026	UNIVERSITY OF MARYLAND, BALTIMORE	220 ARCH ST OFC LEVEL2	BALTIMORE	MD	21201	BALTIMORE CITY	USA	Allergy and Infectious Diseases Research	000	1	9/8/2026	NEW	$769,725
														Subtotal = $769,725

Grand Total All Awards = $769,725

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Culture-Independent Pathogen Identification Through Rapid Lipidomic Profiling

Award Number: R01AI198756

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 09/08/2026

PERIOD OF PERFORMANCE END DATE: 08/31/2031

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