# A "Culture" Shift: Integrated Bacterial Screening and Antibacterial Susceptibility Test on Microfluidic Digital Array for Bloodstream Infections

> **NIH NIH R01** · JOHNS HOPKINS UNIVERSITY · 2021 · $720,032

## Abstract

PROJECT SUMMARY
The ability for clinicians to effectively treat bacterial infections with targeted antibacterials in the acute-care
settings hinges on diagnostics capable of identifying the pathogen broadly and determining its susceptibility to
antibacterials in a timely manner. Bloodstream infection (BSI) is a particularly representative disease because
it is the leading cause of death due to infections with rapid disease progression. Unfortunately, the
inconvenient delay of blood culture for definitive diagnosis contributes to widespread empiric use of broad-
spectrum antibacterials and emergence of multi-drug-resistant pathogens. Toward addressing this critical
unmet need, we propose to develop a new molecular diagnostic platform that integrates bacterial detection,
species identification (ID), and antibacterial susceptibility testing (AST) from blood samples in a streamlined
test. The expected sample-to-answer turnaround time is 90 min for ID and as early as 2-3 hr for AST. Such
integrated diagnostic solution within the proposed timeframe will transform acute-care clinicians’ ability to
establish diagnosis of bacterial infections, need for infection control, and antibacterial treatment based on
objective data to improve clinical outcome.
Using an innovative microfluidic digital array chip for assaying single cells as a backbone technology, we
propose to develop a new molecular diagnostic platform which promises rapid ID and AST and allows
customizable workflow and assay tailored to the clinical scenario while adjustable based on real-time results.
The array chip seamlessly integrates digitization of cells, brief incubation (under various drug conditions),
single-cell PCR (scPCR) or reverse transcriptase PCR (scRT-PCR) and single-cell high-resolution melt
(scHRM). Thereby, bacterial pathogen can be detected at the level of single-cells, identified based on species-
specific melt curves, and their antibacterial susceptibility profile subsequently assessed by measuring changes
in rRNA level as a biosynthetic marker of cell viability. ScPCR/scRT-PCR enables sensitive detection and
absolute quantification of rRNA of individual cells critical to rapid and reliable differentiation between viable and
no-viable cells; while scHRM overcomes a key limitation of bulk HRM to resolve multiple species for
diagnosing polymicrobial infections or discarding contaminations. Since both ID and AST do not rely on culture,
they reduce total turnaround time from days to minutes/hours.
We have assembled a superb team of multi-disciplinary investigators and industry advisors with
complementary expertise and strong track record of team science. We propose the following aims:1) to
develop a streamlined BSI diagnostic protocol for integrated ID and AST; 2) to develop a microfluidic array chip
that enables ID and AST with single-cell resolution; 3) to develop instrument and analysis programs for single-
cell ID and AST; and 4) to demonstrate the single-cell diagnostic pla...

## Key facts

- **NIH application ID:** 10078849
- **Project number:** 5R01AI137272-04
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Tza-Huei Jeff Wang
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $720,032
- **Award type:** 5
- **Project period:** 2018-01-23 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10078849

## Citation

> US National Institutes of Health, RePORTER application 10078849, A "Culture" Shift: Integrated Bacterial Screening and Antibacterial Susceptibility Test on Microfluidic Digital Array for Bloodstream Infections (5R01AI137272-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10078849. Licensed CC0.

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