Portable System for On-Site Detection of Legionella

EPA Contract Number: 68D99085
Title: Portable System for On-Site Detection of Legionella
Investigators: Fernandez, Salvador M.
Small Business: Ciencia Inc.
EPA Contact: Richards, April
Phase: II
Project Period: September 1, 1999 through September 1, 2001
Project Amount: $225,000
RFA: Small Business Innovation Research (SBIR) - Phase II (1999) Recipients Lists
Research Category: Ecological Indicators/Assessment/Restoration , SBIR - Monitoring , Small Business Innovation Research (SBIR)


The objective of the proposed program is to develop a fluorescence-based system (instrument plus reagents) for onsite quantitative detection of Legionella. Currently, Legionella testing must be performed in the laboratory, which is both costly and slow. There is a need for improved methods capable of rapid onsite analysis. The proposed system will be hand-portable, low-cost, and capable of high sensitivity. In addition, it will provide immediate results (approximately 1 hour), and be able to distinguish between live and dead Legionella bacteria. No such systems currently are available. Detection will be based on the integration of an innovative fluorescence immunoassay using phycobilisome markers and compact fluorescence instrumentation recently developed at Ciencia. In Phase I, a conjugation procedure was developed, and conjugates were made that stained Legionella brightly and specifically as demonstrated by microscopic examination of stained bacteria in a fluorescence microscope. Also, a breadboard prototype of a light emitting diode-based fluorimeter was built, and an assay protocol was developed. Results demonstrated that the proposed dye provides more than three orders of magnitude greater sensitivity than fluorescein, the dye currently used in the DFA Legionella assay. In Phase II, an engineering prototype of a portable instrument will be built and field-validated with real-world samples.

Supplemental Keywords:

small business, SBIR, monitoring, analytical, engineering, chemistry, EPA., RFA, Scientific Discipline, Water, Ecosystem Protection/Environmental Exposure & Risk, Monitoring/Modeling, Microbiology, Analytical Chemistry, Engineering, Chemistry, & Physics, environmental monitoring, microbial monitoring, pathogens, field portable monitoring, field portable systems, bacteria monitoring, field monitoring, municipal wastewater, biomonitoring, fluorescence detection, Ligionella detection, indoor air, measurement, innovative technologies, flourescence assay, microbial flora, atmospheric chemistry

Progress and Final Reports:

  • Final Report

  • SBIR Phase I:

    Portable System for On-Site Legionella Detection  | 1999 Progress Report  | Final Report