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J Microbiol Methods. 2015 Nov;118:51-6. doi: 10.1016/j.mimet.2015.08.011. Epub 2015 Aug 23.

Rapid detection of Bacillus anthracis by γ phage amplification and lateral flow immunochromatography.

Author information

1
Colorado School of Mines, Department of Chemistry and Geochemistry, Golden, CO 80401, United States.
2
Romonics, LLC, Boulder, CO 80301, United States.

Abstract

New, rapid point-of-need diagnostic methods for Bacillus anthracis detection can enhance civil and military responses to accidental or deliberate dispersal of anthrax as a biological weapon. Current laboratory-based methods for clinical identification of B. anthracis require 12 to 120h, and are confirmed by plaque assay using the well-characterized γ typing phage, which requires an additional minimum of 24h for bacterial culture. To reduce testing time, the natural specificity of γ phage amplification was investigated in combination with lateral flow immunochromatography (LFI) for rapid, point-of-need B. anthracis detection. Phage-based LFI detection of B. anthracis Sterne was validated over a range of bacterial and phage concentrations with optimal detection achieved in as little as 2h from the onset of amplification with a threshold sensitivity of 2.5×10(4)cfu/mL. The novel use of γ phage amplification detected with a simple, inexpensive LFI assay provides a rapid, sensitive, highly accurate, and field-deployable method for diagnostic ID of B. anthracis in a fraction of the time required by conventional techniques, and without the need for extensive laboratory culture.

KEYWORDS:

Bacillus anthracis; Bacteriophage amplification; Lateral flow immunochromatography; Point-of-need diagnostics

PMID:
26310605
DOI:
10.1016/j.mimet.2015.08.011
[Indexed for MEDLINE]

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