Ferrofluid-based assay methods, and systems for parasite eggs or oocysts detection
AssigneeAncera, LLC
InventorsMary K.H. SMITH, Arjun GANESAN, John C. VOYTA
Life of this patent
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Abstract
Embodiments of the present disclosure are directed to ferrofluid-based assay methods, systems and device for detecting one or more parasite oocyst or egg, and more specifically, to detecting parasite oocysts or eggs in fecal matter of livestock, so as to determine parasitic infections within such livestock. Such embodiments are configured to help in the ability to maintain healthy livestock for human consumption.
Claims72
An assay system configured to at least one of detect, enumerate, and characterize one or more parasite oocyst or egg from at least one fecal sample or a plurality of samples, comprising:
- a ferrofluidic assay device configured to receive a ferrofluidic cartridge;
- the ferrofluidic cartridge including a plurality of windows;
- an imager configured to image each window of the cartridge either separately or together, wherein detecting, enumerating, and characterizing one or more parasite oocyst or egg is determined by an analysis of one or more images;
- a controller configured to control at least one of the ferrofluidic assay device, the ferrofluidic cartridge, and the imager;
- and
- assay processing components comprising at least one of reagents, and controls
- wherein the system is configured to at least one of:
- separate any and all of parasite oocysts or eggs;
- move or otherwise locate the parasite oocysts or eggs to one or more of the windows;
- detect, enumerate/count, and/or characterize the parasite oocysts or eggs.
The system of any of claims 1-4, wherein the ferrofluid includes at least one density modifying agents.
The system of any one of the preceding claims, wherein the system is configured to flow at least one of the plurality of samples into cartridge for a predetermined period of time.
A method for detecting parasitic infection in a sample in an assay system comprising:
- preparing a plurality of fecal samples for analysis, wherein preparing comprises, for each sample:
- labeling at least each of a first tube and a second tube placing a predetermined amount of sample within the first tube;
- placing a predetermined amount of NaOH at a pre-determined concentration in the first tube;
- vortexing the first tube for a first predetermined period of time;
- placing a predetermined amount of Sheather's sugar solution to the first tube;
- vortexing the first tube for a second predetermined period of time;
- optionally removing an amount of the sample from the first tube and determining a reference count of oocysts;
- transferring a third predetermined amount of each fecal-NaOH mixture of each first tube into a respective second tube;
- adding a fourth predetermined amount of ferrofluid and a fifth predetermined amount of SYBR into each respective second tube;
- vortexing each respective second tube for a third predetermined amount of time
- add the vortexed samples to lanes of an assay cartridge having a plurality of channels/lanes, each channel configured with at least one capture region/zone having at least one unique agent for binding with at least one specific parasite oocyst or egg;
- flowing the mixture over at least one of the channels of the cartridge, wherein electromagnetic fields produced by the system and acting on the ferrofluid push up oocysts contained in the at least one sample to a capture region proximate a channel window provided for each channel;
- scanning each window of each channel to produce a respective scan image thereof;
- processing each respective scan image so as to at least one of identify and count each oocyst imaged from a respective channel window;
- converting each count from each processed image to oocysts per gram of original fecal sample;
- and
- optionally identifying and measuring the major axis of each counted oocyst such that the number of small (<18 μm), medium (18-25 μm), and large (>25 μm) oocysts are counted for each sample and optionally converted to oocysts per gram of original fecal sample.
A method for detecting parasitic infection in fecal samples in an assay system comprising:
- optionally placing a predetermined amount of animal fecal matter into a first side (“fecal side”) of at least a two-sided sample filter bag (“sample bag”);
- adding a predetermined amount of NaOH to the first side of the sample bag forming a first mixture;
- performing a first homogenization of the mixture, via, for example, massaging the mixture for a first predetermined period of time;
- incubating the mixture for a second predetermined period of time;
- adding in a predetermined amount of Sheather's solution to a second side of the sample bag (“filtered side”);
- performing a second homogenization of the mixture, via, for example, massaging the mixture of a second predetermined period of time;
- removing an aliquot of a predetermined amount from the filtered side of the bag;
- transferring the aliquot to a tube of predetermined size;
- adding a predetermined amount of ferrofluid to the tube;
- vortexing the tube for a predetermined period of time;
- transferring the contents of the tube to a single sample well of the assay cartridge;
- flowing each sample from each sample well into an imaging window;
- push all cells to the surface of the imaging zone;
- optionally repeating the above-noted steps a plurality of times so as to utilize a corresponding number of sample wells, such that, each sample corresponds to a unique sample well,
- optionally flowing each sample from each sample well into at least one imaging/capture zone of the assay cartridge so as to capture at least one specific parasite oocyst or egg contained in the sample;
- acquiring image of each imaging zone;
- determining a number of parasite oocysts or eggs present in the imaging zone;
- and
- determining a size distribution of the parasite oocysts or eggs captured in the at least one imaging zone.
The method, system or device of any one of the preceding claims, further configured to assess a size distribution of Eimeria oocysts in vaccine preparation.
Dependencies are inferred from the claim text.