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Title: Universal Bacterial Biosensor

Abstract

This report describes methods for rapid differentiation of bacterial pathogens directly in patient blood to inform treatment.

Authors:
 [1]
  1. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Publication Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
USDOE National Nuclear Security Administration (NNSA)
OSTI Identifier:
1369153
Report Number(s):
LA-UR-17-25391
DOE Contract Number:
AC52-06NA25396
Resource Type:
Technical Report
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES

Citation Formats

Kubicek-Sutherland, Jessica Zofie. Universal Bacterial Biosensor. United States: N. p., 2017. Web. doi:10.2172/1369153.
Kubicek-Sutherland, Jessica Zofie. Universal Bacterial Biosensor. United States. doi:10.2172/1369153.
Kubicek-Sutherland, Jessica Zofie. 2017. "Universal Bacterial Biosensor". United States. doi:10.2172/1369153. https://www.osti.gov/servlets/purl/1369153.
@article{osti_1369153,
title = {Universal Bacterial Biosensor},
author = {Kubicek-Sutherland, Jessica Zofie},
abstractNote = {This report describes methods for rapid differentiation of bacterial pathogens directly in patient blood to inform treatment.},
doi = {10.2172/1369153},
journal = {},
number = ,
volume = ,
place = {United States},
year = 2017,
month = 7
}

Technical Report:

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  • An in vitro method for monitoring cadmium, one of the most lethal bivalent heavy metals, can detect biologically active levels. The effects of cadmium tend to concentrate in protozoa far above natural levels and therein begin transferring through freshwater food chains to animals and humans. In a small sample volume (approximately 5 ml) the method uses the toxic response to the protozoa, Tetrahymena pyriformis, to cadmium. The assay relies on macroscopic bioconvective patterns to measure the toxic response, giving a sensitivity better than 1 micro-g/1 and a toxicity threshold to 7 micro-g/1 for Cd(2+). Cadmium hinders pattern formation in amore » dose-dependent manner. Arrested organism growth arises from slowed division and mutation to non-dividing classes. Unlike previous efforts, this method can be performed in a shallow flow device and does not require electronic or chemical analyses to monitor toxicity.« less
  • A methodology is described for immobilizing the enzyme alkaline phosphatase onto a glass surface using a novel biotinylated copolymer, poly(3-undecylthiophene-co-3- methanoithiophene). A streptavidin conjugate of alkaline phosphatase is used in this study. The biotinylated polymer is attached to the silanized glass surface via hydrophobic interactions and the enzyme is interfaced with the polymer through the classical biotin- streptavidin interaction. Alkaline phosphatase catalyzes the dephosphorylation of a macrocyclic compound, chloro-3-(4-methoxy spiro) (1,2 dioxetane-3-2`-tricyclo-) (3.3.1.1 )-(decani-4-yl) phenyl phosphate, to a species which emits energy by chemiluminescence. This chemiluminescence signal can be detected with a photomultiplier tube for enzymatic catalysis with the biocatalystmore » both in solution and immobilized on a glass surface. The signal generation is inhibited by the organophosphorus based insecticides such as paraoxon as well as nerve agents. We demonstrate in this study that a number of organophosphorus based insecticides inhibit the enzyme-mediated generation of chemiluminescence signal. This is true for the enzyme conjugate both free in solution and immobilized on a glass surface. In solution, the inhibition resembles the case of a partially uncompetitive system. By this type of inhibition we are able to detect pesticides down to about 50 ppb for the enzyme in solution. The pesticide detection limit of immobilized enzyme is currently being investigated. The enzyme is capable of a number of measurement cycles without significant loss of signal level.« less
  • The purpose of this contract was to explore and demonstrate the application of technology based on the Light Addressable Potentiometric Sensor (LAPS) to detection and verification problems for bio-chemical warfare agents. The principal analytical method employed was microphysiometry, by which perturbations of cell physiology are detected using the LAPS. The work was done in three principal segments. First, silicon microtechnology was used to design and fabricate sensor chips and microfluidic components suitable for the project. Second, a systems-engineering effort designed and assembled a prototype high-performance microphysiometer. Third, a biological effort employed a wide variety of cellular and ligand-receptor model systemsmore » to validate the generality of the method and the performance of the prototype instrument.« less
  • The purpose of this contract was to explore and demonstrate the application of technology based on the Light Addressable Potentiometric Sensor (LAPS) to detection and verification problems for bio-chemical warfare agents. The principal analytical method employed was microphysiometry, by which perturbations of cell physiology are detected using the LAPS. The work was done in three principal segments. First, silicon microtechnology was used to design and fabricate sensor chips and microfluidic components suitable for the project. Second, a systems-engineering effort designed and assembled a prototype high-performance microphysiometer. Third, a biological effort employed a wide variety of cellular and ligand-receptor model systemsmore » to validate the generality of the method and the performance of the prototype instrument.« less