Methods for performing determinations of immune reactants in biological fluids
First Claim
1. A method for performing in vitro determinations of first immune reactants in biological fluids of humans or animals, comprising the steps of:
- (a) permanently coloring solid substrate material of a plurality of test units to render each test unit visually discernable from the other test units by its color, and further including the steps of;
(a.1) forming each of said test units as an elongate rod having a known diameter and a test tip affixed to a distal end of the rod, said test tip being formed of said substrate material to diverge from the distal end of the rod to a maximum transverse dimension greater than said known diameter; and
(a.2) permanently coloring different test unit rods and test tips with different colors to render the combination of the rod color and tip color for each test unit representative of the identity of at least one second immune reactant adsorbed on said each test unit;
(b) adsorbing on the substrate of each test unit at least one of plural second immune reactants such that the second immune reactant adsorbed on each test unit is identified by the color of the substrate material of that test unit;
(c) suspending the plural test units in a test array;
(d) inserting the array of plural second immune reactant-coated and permanently colored test units into the biological fluid disposed in respective reaction containers to permit any first immune reactants in the biological fluid that are specific to a particular second immune reactant coating on any of the test units to become bound to the particular second immune reactant on said any test unit;
(e) removing said test units from said biological fluid and then washing and drying the test units;
(f) incubating the plurality of permanently colored test units, with said first immune reactants bound thereto as in step (d), in an enzyme-labeled conjugate liquid in respective reaction containers to cause the conjugate liquid to react with the first immune reactants bound to the plurality of permanently colored test units;
(g) removing said test units from said conjugate liquid and then washing and drying said test units;
(h) incubating the plurality of permanently colored test units in chromogenic substrate liquid in respective containers to develop specific color in the chromogenic liquid dependent upon the first immune reactants that were bound to the test units in step (d);
(i) removing the test units from the chromogenic substrate liquid; and
(j) determining concentrations of said first immune reactants in the biological fluid into which said test units were inserted in step (b) by analyzing the remaining chromogenic substrate liquid from respective containers for color development and intensity.
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Abstract
Apparatus for performing determinations of immune reactants (e.g., antigens, antibodies) in bodily fluids includes multiple test units having respective elongated rods with transversely-expanded tips at their distal ends. The tips are each coated with respective immune reactants (e.g., allergens) which react in a known manner with respective allergen-specific or allergen-binding antibodies in human serum. The test units are color-coded to identify the allergen coatings and are supported at their proximal ends and positionally identified on a strip. By correlating the color code to a chart, the specific immune reactant (e.g., allergen) coating can be easily identified. The supporting strip for the test unit has through-holes which frictionally engage the proximal ends of the test unit rods with a spacing that permits all of the supported test units to be simultaneously inserted into an assembly of reaction containers arranged in a linear array. Additionally, the through-hole spacing in the strip permits alternate test units to be simultaneously placed in an array of test tubes that are transversely larger than the reaction containers and supported in a linear array in a test tube rack. The tip of each test unit may take the form of two frusto-conical sections joined by a short cylindrical section extending longitudinally between their larger ends. Alternatively, the frusto-conical sections may be multi-faceted to minimize scraping of the allergen coating from the tip during insertion and removal of the test units into the reaction containers/test tubes.
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Citations
6 Claims
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1. A method for performing in vitro determinations of first immune reactants in biological fluids of humans or animals, comprising the steps of:
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(a) permanently coloring solid substrate material of a plurality of test units to render each test unit visually discernable from the other test units by its color, and further including the steps of; (a.1) forming each of said test units as an elongate rod having a known diameter and a test tip affixed to a distal end of the rod, said test tip being formed of said substrate material to diverge from the distal end of the rod to a maximum transverse dimension greater than said known diameter; and (a.2) permanently coloring different test unit rods and test tips with different colors to render the combination of the rod color and tip color for each test unit representative of the identity of at least one second immune reactant adsorbed on said each test unit; (b) adsorbing on the substrate of each test unit at least one of plural second immune reactants such that the second immune reactant adsorbed on each test unit is identified by the color of the substrate material of that test unit; (c) suspending the plural test units in a test array; (d) inserting the array of plural second immune reactant-coated and permanently colored test units into the biological fluid disposed in respective reaction containers to permit any first immune reactants in the biological fluid that are specific to a particular second immune reactant coating on any of the test units to become bound to the particular second immune reactant on said any test unit; (e) removing said test units from said biological fluid and then washing and drying the test units; (f) incubating the plurality of permanently colored test units, with said first immune reactants bound thereto as in step (d), in an enzyme-labeled conjugate liquid in respective reaction containers to cause the conjugate liquid to react with the first immune reactants bound to the plurality of permanently colored test units; (g) removing said test units from said conjugate liquid and then washing and drying said test units; (h) incubating the plurality of permanently colored test units in chromogenic substrate liquid in respective containers to develop specific color in the chromogenic liquid dependent upon the first immune reactants that were bound to the test units in step (d); (i) removing the test units from the chromogenic substrate liquid; and (j) determining concentrations of said first immune reactants in the biological fluid into which said test units were inserted in step (b) by analyzing the remaining chromogenic substrate liquid from respective containers for color development and intensity.
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2. A method for performing in vitro determinations of first immune reactants in biological fluids of humans or animals, comprising the steps of:
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(a) permanently coloring solid substrate material of a plurality of test units to render each test unit visually discernable from the other test units by its color, and further including the step of; (a.1) forming each of said test units as an elongate rod having a known diameter and a test tip affixed to a distal end of the rod, said test tip being formed of said substrate material to diverge from the distal end of the rod to a maximum transverse dimension greater than said known diameter; (b) adsorbing on the substrate of each test unit at least one of plural second immune reactants such that the second immune reactant adsorbed on each test unit is identified by the color of the substrate material of that test unit; (c) suspending the plural test units in a test array and further including the steps of; (c.1) establishing said test array with an elongate holder strip; (c.2) defining a plurality of spaced through-holes in said strip; and (c.3) frictionally engaging a proximal end of each rod in a respective through hole in said strip; (d) inserting the array of plural second immune reactant-coated and permanently colored test units into the biological fluid disposed in respective reaction containers to permit any first immune reactants in the biological fluid that are specific to a particular second immune reactant coating on any of the test units to become bound to the particular second immune reactant on said any test unit; (e) removing said test units from said biological fluid and then washing and drying the test units; (f) incubating the plurality of permanently colored test units, with said first immune reactants bound thereto as in step (d), in an enzyme-labeled conjugate liquid in respective reaction containers to cause the conjugate liquid to react with the first immune reactants bound to the plurality of permanently colored test units; (g) removing said test units from said conjugate liquid and then washing and drying said test units; (h) incubating the plurality of permanently colored test units in chromogenic substrate liquid in respective containers to develop specific color in the chromogenic liquid dependent upon the first immune reactants that were bound to the test units in step (d); (i) removing the test units from the chromogenic substrate liquid; and (j) determining concentrations of said first immune reactants in the biological fluid into which said test units were inserted in step (b) by analyzing the remaining chromogenic substrate liquid from respective containers for color development and intensity. - View Dependent Claims (3)
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4. A method for performing in vitro determinations of allergen-specific IgE antibodies in human serum comprising the steps of:
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(a) permanently coloring solid substrate material of a plurality of test units to render each test unit visually discernible from the other test units by its color, and further including the steps of; (a.1) forming each of said test units as an elongate rod having a known diameter and a test tip affixed to a distal end of the rod, said test tip being formed of said substrate material to diverge from the distal end of the rod to a maximum transverse dimension greater than said known diameter; and (a.2) permanently coloring different test unit rods and test tips with different colors to render the combination of the rod color and tip color for each test unit representative of the identity of at least one allergen adsorbed on said each test unit; (b) coating at least one allergen to a solid phase on the substrate of each test unit such that each allergen coated on each test unit is identified by the color of the substrate material of that test unit; (c) suspending the plural test units in a test array; (d) immersing the test units into human serum to permit said allergens coated on the test units to react with allergen-specific IgE antibodies in the serum and provide specific allergen-bound IgE on said test units; (e) washing away non-specific reactants from said test units; (f) immersing the test units that have been washed in step (e) into a conjugate comprising enzyme-labeled anti-human IgE to cause the conjugate to react with the allergen-bound human IgE on the test units and provide a bound complex on the test units comprising allergen--IgE--enzyme-labeled anti-human IgE on said test units; (g) further washing the test units; (h) immersing the test units that have been washed in step (g) into a liquid chromogenic substrate specific to said conjugate to cause the substrate to react with the bound complex on said test units; (i) removing the test units from the chromogenic substrate; and (j) spectrophotometrically determining the amount of circulating allergen-specific IgE antibodies in the human serum as a function of the color intensity of the chromogenic substrate.
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5. A method for performing in vitro determinations of allergen-specific IgE antibodies in human serum comprising the steps of:
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(a) permanently coloring solid substrate material of a plurality of test units to render each test unit visually discernible from the other test units by its color, and further including the step of; (a.1) forming each of said test units as an elongate rod having a known diameter and a test tip affixed to a distal end of the rod, said test tip being formed of said substrate material to diverge from the distal end of the rod to a maximum transverse dimension greater than said known diameter; (b) coating at least one allergen to a solid phase on the substrate of each test unit such that each allergen coated on each test unit is identified by the color of the substrate material of that test unit; (c) suspending the plural test units in a test array, and further including the steps of; (c.1) establishing said test array with an elongate holder strip; (c.2) defining a plurality of spaced through-holes in said strip; and (c.3) frictionally engaging a proximal end of each rod in a respective through hole in said strip; (d) immersing the test units into human serum to permit said allergens coated on the test units to react with allergen-specific IgE antibodies in the serum and provide specific allergen-bound IgE on said test units; (e) washing away non-specific reactants from said test units; (f) immersing the test units that have been washed in step (e) into a conjugate comprising enzyme-labeled anti-human IgE to cause the conjugate to react with the allergen-bound human IgE on the test units and provide a bound complex on the test units comprising allergen--IgE--enzyme-labeled anti-human IgE on said test units; (g) further washing the test units; (h) immersing the test units that have been washed in step (g)into a liquid chromogenic substrate specific to said conjugate to cause the substrate to react with the bound complex on said test units; (i) removing the test units from the chromogenic substrate; and (j) spectrophotometrically determining the amount of circulating allergen-specific IgE antibodies in the human serum as a function of the color intensity of the chromogenic substrate. - View Dependent Claims (6)
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Specification