Inexpensive High-Throughput Multiplexed Cytokine Detection for Tuberculosis Diagnostics Using Amplified Enzymatic...

Area of Science:

• Biomedical Diagnostics
• Point-of-Care Testing
• Biotechnology

Background:

• Accurate and affordable diagnostics are crucial for tuberculosis (TB) control, especially in resource-poor endemic settings.
• Current Interferon-gamma Release Assays (IGRAs) for latent TB infection (LTBI) require specialized equipment, limiting their use in low-resource areas.
• Multiplexed cytokine detection can enhance diagnostic accuracy but necessitates expensive instrumentation, with no current point-of-care solutions available.

Purpose of the Study:

• To develop and validate a low-cost, high-throughput, multiplexed cytokine detection platform for TB diagnostics.
• To enable sensitive detection of multiple cytokines, including Interferon-gamma (IFN-γ), Interleukin-2 (IL-2), and Tumor Necrosis Factor alpha (TNF-α).
• To create a point-of-care compatible assay that utilizes common materials and cellphone-based readout.

Main Methods:

• Development of a multiplexed cytokine detection assay using amplified enzymatic silver metallization on a plastic substrate.
• Assay performed in microwells on a standard plastic petri dish.
• Dry readout achieved using a cellphone camera for cost-effective and simplified data acquisition.

Main Results:

• Demonstrated detection of clinically relevant sub-picomolar levels of multiple cytokines (IFN-γ, IL-2, TNF-α) from small blood volumes (<5µL).
• Successfully distinguished between IGRA-positive and IGRA-negative participant samples from a TB-endemic setting.
• The platform significantly reduces cost and complexity compared to existing multiplexed cytokine detection methods.

Conclusions:

• The developed platform offers a low-cost, high-throughput, and point-of-care compatible solution for multiplexed cytokine detection.
• This technology has the potential to overcome critical diagnostic bottlenecks in TB control and elimination efforts globally.
• The cellphone-based readout system simplifies complex assays, making advanced diagnostics more accessible in resource-limited environments.