Detection of Bacterial and Viral Pathogens Using Fluorescence Activated Sensing Technology (FAST System)

Categories: “Diagnostics” “Viruses, Chronic & Infectious Diseases” “Research Tools“

Reference #: 2004-002 & 2009-010

OTC Contact: Ruchika Nijhara Ph.D., MBA (Directory Information | Send a Message)

Description

Researchers at Georgetown University have developed a sensitive method for detecting the presence of any microorganism (such as bacteria or virus) in an infection, contamination or otherwise that affect human and animal health. The method involves a real time isothermal reaction that amplifies the signal generated by a fluorescent DNA probe that binds specifically to the target organism. In cases where maximum speed and sensitivity are required, signal amplification occurs at the same time as targeted DNA amplification.

Applications

• Diagnosis of infectious diseases (human, animal or plant diseases)
• Diagnosis of hospital infections, such as MRSA
• Diagnosis of COVID-19
• Diagnosis of ESKAPE organisms
• Detection of biowarfare agents- can be used for a rapid, specific, and sensitive detection of any biological threat agent such as anthrax or plaque.
• Food and water surveillance
• Environmental monitoring
• SNP analysis and genotyping
• As a biological research tool

Advantages

• Broad spectrum: detects bacteria and both DNA and RNA viruses
• Sensitive: detection levels approaching a single organism
• Specific: no cross-reactivity with closely related organisms
• Rapid: results in minutes
• Robust: works with unpurified samples
• High throughput: can be multiplexed and automated
• Stability: processed samples can be archived for future testing
• Isothermal: does not use thermocycling
• Multiple applications: works with environmental, biological and diagnostic samples
• Extremely low false positive and false negatives rates
• Extreme sensitivity towards RNA based microorganisms
• No highly specialized equipment is required
• No unusual training is required

Stage of Development

Studies have demonstrated that the assay can be used to specifically detect the presence of (1) Bacillus anthracis pX01 and pX02 plasmids; (2) Escherichia coli genomic DNA; (3) Bacillus subtilis genomic DNA; (4) different Dengue serotypes in a highly sensitive single-tube multiplex assay; (5) Chlamydia trachomatis in human urine.

The initial work was funded by the Department of Defense (DoD) and focused on the development of a stand-alone instrument capable of detecting bioterror threat organisms. As a part of that work, Georgetown scientists developed probes against Bacillus anthracis that have been validated by DoD. The researchers developed and delivered to DoD a semi-automated prototype of a stand-alone detector that uses these probes.

Inventors

Mark Danielsen, Eugene Davidson, Kenneth L. Dretchen, Berenice Alfonso, and Bolor Tumurpurev

Relevant Publications

Sequence specific detection of DNA using nicking endonuclease signal amplification (NESA); Danielsen M et al; Nucleic Acids Res. 2007;35(18):e117.

Patent Status

US issued patents:
9,012,142 issued April 21, 2015
9,562,258 issued Feb, 7, 2017
10,023,905 issued July 17, 2018
10, 246,739 issued April 2, 2019