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US 10,094,829 B2Granted
Method for the quantification of parasite eggs in feces
AssigneeMEP Equine Solutions LLC
InventorsPawel Slusarewicz, Eric W. Hauck
Life of this patent
Estimating the term…
Abstract
Method and kits are provided determining the presence or absence of parasitic helminth eggs in environmental samples, particularly fecal samples. The methods incorporate egg capture methods and the use of N-acetyl-D-glucosamine specific ligands for egg detection.
Claims30
A method of detecting the presence or absence of helminth eggs or protozoan oocysts in an environmental sample, the method comprising, sequentially:
- flowing a solution comprising an environmental sample suspended in water or a sample buffer through a filtration membrane having a pore size of between about 5 microns and about 45 microns to physically capture helminth eggs or protozoan oocysts on the filtration membrane, wherein physically capturing the eggs or oocysts on the filtration membrane occurs in the absence of affinity ligands;
- contacting helminth eggs or protozoan oocysts physically captured on the filtration membrane with a chitin exposing reagent selected from a surfactant, an oxidizing agent, a chaotrope, an enzyme, and bleach;
- contacting helminth eggs or protozoan oocysts physically captured on the filtration membrane with a N-acetyl-D-glucosamine binding protein or fragment thereof conjugated to a detectable moiety;
- and
- imaging the sample physically captured on the filtration membrane using an imaging device appropriate for visualizing the detectable moiety to produce an image of particles comprising the detectable moiety;
- electronically parsing the particles in the image by size, shape, or a combination thereof;
- and
- detecting the presence or absence of helminth eggs or protozoan oocysts in the environmental sample captured on the filtration membrane based on the electronic parsing of the particles in the image.
5 more elements
A method of detecting the presence or absence of helminth eggs or protozoan oocysts in an environmental sample, the method comprising, sequentially:
- flowing a solution comprising an environmental sample suspended in water or a sample buffer through a filtration membrane having a pore size of between about 5 microns and about 45 microns to physically capture helminth eggs or protozoan oocysts on the filtration membrane, wherein physically capturing the eggs or oocysts on the filtration membrane occurs in the absence of affinity ligands;
- contacting helminth eggs or protozoan oocysts physically captured on the filtration membrane with a chitin exposing reagent selected from a surfactant, an oxidizing agent, a chaotrope, an enzyme, and bleach;
- contacting helminth eggs or protozoan oocysts physically captured on the filtration membrane with a chitin binding domain (CBD) protein or fragment thereof conjugated to a detectable moiety;
- and
- imaging the sample physically captured on the filtration membrane using an imaging device appropriate for visualizing the detectable moiety to produce an image of particles comprising the detectable moiety;
- electronically parsing the particles in the image by size, shape, or a combination thereof;
- and
- detecting the presence or absence of helminth eggs or protozoan oocysts in the environmental sample captured on the filtration membrane based on the electronic parsing of the particles in the image;
- and
- wherein the detecting step is performed without removing the helminth eggs or protozoan oocysts physically captured on the filtration membrane.
7 more elements
A method of detecting the presence or absence of helminth eggs or protozoan oocysts in an environmental sample, the method comprising, sequentially:
- flowing a solution comprising an environmental sample suspended in water or a sample buffer through a filtration membrane having a pore size of between about 5 microns and about 45 microns to physically capture helminth eggs or protozoan oocysts on the filtration membrane, wherein the filtration membrane does not contain specific chemicals to bind the eggs or oocysts;
- contacting helminth eggs or protozoan oocysts physically captured on the filtration membrane with a chitin exposing reagent selected from a surfactant, an oxidizing agent, a chaotrope, an enzyme, and bleach;
- contacting helminth eggs or protozoan oocysts physically captured on the filtration membrane with a N-acetyl-D-glucosamine binding protein or fragment thereof conjugated to a detectable moiety;
- and
- imaging the sample physically captured on the filtration membrane using an imaging device appropriate for visualizing the detectable moiety to produce an image of particles comprising the detectable moiety;
- electronically parsing the particles in the image by size, shape, or a combination thereof;
- and
- detecting the presence or absence of helminth eggs or protozoan oocysts in the environmental sample captured on the filtration membrane based on the electronic parsing of the particles in the image.
5 more elements
Dependencies are inferred from the claim text.