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A highly multiplexed broad pathogen detection assay for infectious disease diagnostics.

Jeffrey W Koehler1, Christina E Douglas1, Timothy D Minogue1

  • 1Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland, United States of America.

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Summary

A new highly multiplexed assay rapidly detects 164 pathogens, including viruses and bacteria, aiding infectious disease diagnostics. This tool enhances biosurveillance by providing quick, broad pathogen screening for febrile illnesses.

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Area of Science:

  • Infectious Disease Diagnostics
  • Molecular Biology
  • Biosurveillance

Background:

  • Rapid identification of pathogens is crucial for managing acute febrile illnesses and preventing disease spread.
  • Current diagnostics like PCR and ELISA are effective for known pathogens but may miss novel or geographically specific ones.
  • Metagenomic sequencing offers broad detection but is time-consuming and not point-of-care.

Purpose of the Study:

  • To develop and evaluate a highly multiplexed assay for simultaneous detection of a wide range of human pathogens.
  • To bridge the diagnostic gap between rapid point-of-care tests and comprehensive sequencing methods.
  • To create a tool for improved infectious disease diagnostics and global biosurveillance.

Main Methods:

  • Development of a highly multiplexed assay targeting 164 viruses, bacteria, and parasites.
  • Utilized the NanoString nCounter platform for assay detection.
  • Included high-consequence, endemic, and less prevalent pathogens for broad coverage.

Main Results:

  • The assay successfully detected 113 out of 126 available organisms, representing 98 distinct human pathogen types.
  • Achieved positive detection of critical pathogens like Ebola virus, Lassa virus, Dengue virus serotypes 1-4, Chikungunya virus, yellow fever virus, and Plasmodium falciparum.
  • Demonstrated the assay's capability to identify a broad spectrum of infectious agents.

Conclusions:

  • The developed highly multiplexed assay serves as a rapid, user-friendly screening tool for infectious diseases.
  • This assay has the potential to significantly improve diagnostic capabilities and biosurveillance efforts worldwide.
  • It offers a valuable solution for identifying known and potentially unknown pathogens during febrile illnesses.