# Facile Method for Enhanced Pathogen Detection

> **NIH NIH R43** · INSO BIOSCIENCES INC · 2021 · $250,894

## Abstract

Abstract
Leveraging microfluidic technologies for high-efficiency and high-precision cellular sample processing, Tico
Biosciences is developing a culture-free direct-from-sputum method for high-efficiency host cell and host
genomic DNA (gDNA) removal from clinical tuberculosis (TB) samples, enabling rapid, facile, and cost-effective
whole-genome sequencing (WGS) for TB drug susceptibility diagnostics. Drug-resistant TB is a primary threat
to global TB control, and strategies for treatment and medical intervention are heavily dependent on effective
diagnosis and screening. However, current methods for sample processing and enrichment of Mycobacterium
tuberculosis (Mtb) are severely inadequate. TB diagnostic methods currently involve either WGS directly from
sputum samples, resulting in the overwhelming contamination of host genetic material, or culturing of patient
samples, which is time consuming and leads to delayed patient treatment courses and loss of Mtb heterogeneity.
Available strategies for host cell and host DNA removal are labor-intensive, have high skill barriers, and result in
loss of up to 60% of the initial bacillary load. Consequently, the current state of the art results in poor patient
outcomes, especially for those with mixed or drug resistant strains. To address these limitations, Tico
Biosciences has licensed microfluidic technologies that allow for high-efficiency and high-precision cellular
sample processing in order to remove host cell and gDNA in culture-free direct-from-sputum samples. This
method will significantly reduce (1) total analysis turn-around time, (2) amount of sample handling, (3) user skill
requirements, (4) resource requirements (e.g. laboratory space, electrical power, standard biology lab
equipment, etc.), and (5) per-sample costs for reagents and sequencing by a factor of ten. This method results
in the rapid removal of over 99% of mammalian DNA while retaining 85% of the initial bacillary load from a mixed
sample of spiked human and E. coli cells. In this Phase I project, Tico plans to further optimize the performance
of this microfluidic system with mock sputum and demonstrate the feasibility of this approach on pre-banked TB
human clinical samples. This will be accomplished by: 1) Using Tico's system to recover over 95% of bacteria
directly from sputum samples while removing over 99% of mammalian cells as quantified by fluorescence
microscopy and qPCR; and 2) Demonstrating Tico's system to enable culture-free WGS of Mtb directly from
sputum with fewer than 10% of total reads mapped to the human genome. This project will lead to an overall
faster, easier-to-use, and cheaper method of processing clinical sputum samples for genetic analyses such as
WGS or qPCR and will translate to increased adoption of otherwise prohibitively expensive approaches for early
drug resistance screening, which is critically needed in many regions around the world.

## Key facts

- **NIH application ID:** 10155279
- **Project number:** 1R43AI157563-01
- **Recipient organization:** INSO BIOSCIENCES INC
- **Principal Investigator:** Harvey Chen Tian
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $250,894
- **Award type:** 1
- **Project period:** 2021-06-15 → 2023-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10155279

## Citation

> US National Institutes of Health, RePORTER application 10155279, Facile Method for Enhanced Pathogen Detection (1R43AI157563-01). Retrieved via AI Analytics 2026-07-10 from https://api.ai-analytics.org/grant/nih/10155279. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*