Dual Fluorescence Assay For Determining Viability Of Parasitic Or Non-Parasitic Worms
First Claim
1. A method for evaluating the viability of parasitic or non-parasitic worms, for example helminths, comprising(a) providing a sample comprising one or more parasitic or non-parasitic worms, for example helminths;
- (b) contacting the parasitic or non-parasitic worms with a concentration of a first fluorescent dye sufficient to yield a detectable first colour fluorescence in any live parasitic or non-parasitic worms present in the sample and a concentration of a second fluorescent dye sufficient to yield a detectable second colour fluorescence in any dead parasitic or non-parasitic worms present in the sample; and
(c) detecting the first and second fluorescence of the parasitic or non-parasitic worms.
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Accused Products
Abstract
A two colour fluorescent assay and a kit are described that permit the determination of live and dead parasitic or non-parasitic worms, for example helminths. The assay permits screening for the effect of one or more agents or an event on the viability of a parasitic or non-parasitic worm. In one example, the assay comprises (a) providing a sample comprising one or more parasitic or non-parasitic worms; (b) contacting the parasitic or non-parasitic worms with a concentration of fluorescein diacetate sufficient to yield detectable green fluorescence in any live parasitic or non-parasitic worms present in the sample and a concentration of propidium iodide sufficient to yield detectable red fluorescence in any dead parasitic or non-parasitic worms present in the sample; and (c) detecting the red and green fluorescence of the parasitic or non-parasitic worms. The assay further comprises contacting the parasitic or non-parasitic worms with one or more test agents, or subjecting the parasitic or non-parasitic worms to an event. The assay allows for a rapid and objective score of parasitic or non-parasitic worm death and survival and enables the performance of high- throughput screens for the identification of potential agents against diseases such as schistosomiasis. Also described are methods and kits for discriminatory analysis of parasitic or non-parasitic worm phenotype and/or metabolics using FTIR analysis.
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Citations
39 Claims
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1. A method for evaluating the viability of parasitic or non-parasitic worms, for example helminths, comprising
(a) providing a sample comprising one or more parasitic or non-parasitic worms, for example helminths; -
(b) contacting the parasitic or non-parasitic worms with a concentration of a first fluorescent dye sufficient to yield a detectable first colour fluorescence in any live parasitic or non-parasitic worms present in the sample and a concentration of a second fluorescent dye sufficient to yield a detectable second colour fluorescence in any dead parasitic or non-parasitic worms present in the sample; and (c) detecting the first and second fluorescence of the parasitic or non-parasitic worms. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28)
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27. A method for detecting phenotypic and/or metabolic changes in a sample of living parasitic or non-parasitic worms, for example helminths following contact with one or more test agents or after subjecting the sample to an event, the method comprising
(a) providing a sample comprising one or more parasitic or non-parasitic worms, for example helminths; -
(b) contacting the parasitic or non-parasitic worms with one or more test agents or subjecting the sample to an event; (c) performing FTIR spectral analysis of the sample; and (d) comparing the spectra produced from the sample with an FTIR spectra from one or more reference standards indicative of abnormal phenotype and/or metabolic change to identify the presence of one or more abnormal phenotypes and/or metabolic changes in the sample.
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29. A kit for evaluating the viability of parasitic or non-parasitic worms, for example helminths, the kit comprising:
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(i) a microtiter plate comprising a plurality of wells; (ii) a culture media; (iii) a first fluorescent dye that yields a detectable first colour fluorescence in any live parasitic or non-parasitic worms; and (iv) a second fluorescent dye that yields a detectable second colour fluorescence in any dead parasitic or non-parasitic worms. - View Dependent Claims (30, 31, 32, 33, 34, 35, 36, 37, 38)
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39. A kit for evaluating phenotypic and/or metabolic changes of parasitic or non-parasitic worms, for example helminths following contact with one or more test agents or after subjecting a sample of parasitic or non-parasitic worms, for example helminths to an event, the kit comprising:
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(i) a microtiter plate comprising a plurality of wells; (ii) a culture media; and (iii) instructions for performing FTIR analysis.
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Specification