Method and apparatus for coagulation based assays
First Claim
1. A method of performing a coagulation-based assay on a fresh whole blood sample, comprising the steps of:
- providing a cuvette having at least one substantially non-capillary conduit having a first end and a second end, said at least one conduit having a restriction disposed between said first end and said second end of said conduit;
introducing a predetermined volume of said blood sample into said first end of said at least one conduit;
providing an anticoagulating agent at a first position in said at least one conduit, wherein said anticoagulating agent is capable of mixing with said blood sample;
providing a coagulation agent in a second position in said at least one conduit, a portion of said conduit between said first position and said second position curving in a half-circle formation, wherein said coagulation reagent is capable of mixing with said blood sample;
providing a recalcification agent at a third position in said at least one conduit, said third position located between said restriction and said second end of said at least one conduit, wherein said recalcification agent is capable of mixing with said blood sample;
moving said blood sample from said first position to said second position, and from said second position to said third position; and
measuring the time it takes for said blood sample to clot.
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Accused Products
Abstract
An activated partial thromboplastin time (APTT) test is described which employs fresh, whole blood that has not been anticoagulated. The method employs a disposable cuvette having specific reagents placed therein. A sample of whole blood is introduced into the cuvette which is drawn into at least one conduit by a pneumatic pump. Initially, the calcium in the sample is chelated at a first reaction site in the conduit, and thereafter is moved through the conduit to a second reaction site where the sample is coagulated by an APTT reagent. After a predetermined incubation time, the sample is moved to a third reaction site where it is recalcified. Clotting time is measured from the time that the sample is recalcified to the time that clot formation is visibly detected.
112 Citations
23 Claims
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1. A method of performing a coagulation-based assay on a fresh whole blood sample, comprising the steps of:
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providing a cuvette having at least one substantially non-capillary conduit having a first end and a second end, said at least one conduit having a restriction disposed between said first end and said second end of said conduit;
introducing a predetermined volume of said blood sample into said first end of said at least one conduit;
providing an anticoagulating agent at a first position in said at least one conduit, wherein said anticoagulating agent is capable of mixing with said blood sample;
providing a coagulation agent in a second position in said at least one conduit, a portion of said conduit between said first position and said second position curving in a half-circle formation, wherein said coagulation reagent is capable of mixing with said blood sample;
providing a recalcification agent at a third position in said at least one conduit, said third position located between said restriction and said second end of said at least one conduit, wherein said recalcification agent is capable of mixing with said blood sample;
moving said blood sample from said first position to said second position, and from said second position to said third position; and
measuring the time it takes for said blood sample to clot. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A method of performing a coagulation-based assay on a fresh whole blood sample, comprising the steps of:
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providing a cuvette having at least one substantially non-capillary conduit, said at least one conduit having a first end and a second end, wherein at least one restriction is disposed between said first end and said second end of said conduit;
introducing a predetermined volume of said blood sample into said first end of said at least one conduit;
providing a calcium chelating agent at a first position in said at least one conduit in a predetermined amount, wherein said calcium chelating agent mixes with said blood sample for a predetermined period of time;
moving said calcium chelated blood sample along said at least one conduit to a second position, a portion of said conduit between said first position and said second position curving in a half-circle formation;
providing a coagulation agent at said second position in said at least one conduit in a predetermined amount, wherein said coagulation agent mixes with said calcium chelated blood sample for a predetermined period of time;
moving said coagulated blood sample along said at least one conduit to a third position;
providing a recalcification agent at said third position in said at least one conduit in a predetermined amount, said third position located between said at least one restriction and said second end of said at least one conduit, wherein said coagulated blood sample contacts said recalcification agent; and
measuring the time it takes for said blood sample to clot. - View Dependent Claims (9, 10, 11, 12, 13, 14, 15, 16, 21)
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17. A method of performing an activated partial thromboplastin time test on a fresh whole blood sample, comprising the steps of:
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providing a cuvette having at least one substantially non-capillary conduit having a first end and a second end, wherein at least one restriction is disposed between said first end and said second end of said at least one conduit;
introducing a predetermined volume of said blood sample into said first end of said at least one conduit of said cuvette;
providing an anticoagulating agent at a first position in said at least one conduit, wherein said anticoagulating agent mixes with said blood sample for a predetermined period of time to chelate calcium present in said blood sample;
moving said anticoagulated blood sample along said at least one conduit to a second position, a portion of said conduit between said first position and said second position curving in a half-circle formation;
providing a coagulation agent at said second position in said at least one conduit, wherein said coagulation agent mixes with said anticoagulated blood sample for a predetermined period of time to allow activation of deficiency factors and coagulation of said blood sample;
providing a recalcification agent at a third position in said at least one conduit, said third position located between said at least one restriction and said second end of said at least one conduit;
moving said coagulated blood sample through said at least one restriction to said third position, wherein said coagulated blood sample contacts and is recalcified by said recalcification agent; and
measuring the time it takes for said blood sample to clot from the time the coagulated blood sample contacts said recalcification agent to the time that said at least one restriction in said cuvette is blocked by a clot formation. - View Dependent Claims (18, 19, 20, 22, 23)
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Specification