Methods for detecting and measuring aggregation

Abstract

Methods, compositions, systems, and devices are provided for performing and analyzing agglutination assays. In one aspect, methods for image analysis of agglutination assays are provided. In another aspects, methods for performing agglutination assays are provided. In one aspect, the methods may be used for the detection of various molecules, including viruses or antibodies against a virus. In another aspect, the methods can be used to determine effective immunization of a subject.

1 - 29 . (canceled) 
     
     
         30 . A method for determining the presence of an antibody in a biological sample, wherein said antibody binds selectively to a viral particle, the method comprising:
 (a) incubating a mixture of erythrocytes, the viral particle, and the biological sample suspected of containing said antibody, under conditions permitting agglutination of the erythrocytes via interaction with said viral particle;   (b) concurrent with or subsequent to (a), providing the mixture in a vessel having an opening at the top of the vessel and interior conical shape at the bottom of the vessel;   (c) tilting the vessel such that after tilting, at least a portion of the mixture is closer to the opening at the top of the vessel than prior to the tilting, and wherein at least a portion of the sample remains in the vessel after the tilting; and   (d) capturing with the aid of an optical device an image of said mixture, wherein the position of erythrocytes within the tilted vessel may be used to determine the presence or amount of said antibody in the sample.   
     
     
         31 . (canceled) 
     
     
         32 . The method of  claim 30 , wherein one or more zones for analysis are present or established within the vessel, and wherein optical signal is measured over each zone. 
     
     
         33 . The method of  claim 32 , wherein the measured optical signal over the one or more zones may be used to determine the amount of agglutinated or unagglutinated erythrocytes in the sample. 
     
     
         34 . The method of  claim 30 , wherein an axis for analysis is present or established within the vessel, and wherein optical signal is measured along the axis. 
     
     
         35 . The method of  claim 34 , wherein the axis is through the center of a button portion and, if present, a teardrop portion of the packed red cells at the bottom of the vessel. 
     
     
         36 . The method of  claim 34 , wherein the measured optical signal along the axis may be used to determine the amount of agglutinated or unagglutinated erythrocytes in the sample. 
     
     
         37 . The method of  claim 30 , wherein packed red cells at the bottom of the vessel are identified, and the area or perimeter of the packed red cells is measured. 
     
     
         38 . The method of  claim 37 , wherein the method further comprises using a pattern recognition method to identify a button and, optionally, teardrop region of the packed red cells. 
     
     
         39 . The method of  claim 37 , wherein the area or perimeter of the packed red cells may be used to determine the amount of agglutinated or unagglutinated erythrocytes in the sample. 
     
     
         40 - 66 . (canceled) 
     
     
         67 . The method of  claim 30 , wherein said image is a microscopic image. 
     
     
         68 . A method for determining the presence of an antibody in a biological sample, wherein said antibody binds selectively to an agglutinating particle, the method comprising:
 (a) incubating a mixture of visualization particles, the agglutinating particle, and the biological sample suspected of containing said antibody, under conditions permitting agglutination of the visualization particle via interaction with said agglutinating particle;   (b) concurrent with or subsequent to (a), providing the mixture in a vessel having an opening at the top of the vessel and interior conical shape at the bottom of the vessel;   (c) tilting the vessel such that after tilting, at least a portion of the mixture is closer to the opening at the top of the vessel than prior to the tilting, and wherein at least a portion of the sample remains in the vessel after the tilting; and   (d) capturing with the aid of an optical device an image of said mixture, wherein the position of visualization particles within the tilted vessel may be used to determine the presence or amount of said antibody in the sample.   
     
     
         69 . (canceled) 
     
     
         70 . The method of  claim 68 , wherein one or more zones for analysis are present or established within the vessel, and wherein optical signal is measured over each zone. 
     
     
         71 . The method of  claim 70 , wherein the measured optical signal over the one or more zones may be used to determine the amount of agglutinated or unagglutinated visualization particles in the sample. 
     
     
         72 . The method of  claim 68 , wherein an axis for analysis is present or established within the vessel, and wherein optical signal is measured along the axis. 
     
     
         73 . The method of  claim 72 , wherein the axis is through the center of a button portion and, if present, a teardrop portion of the packed visualization particles at the bottom of the vessel. 
     
     
         74 . The method of  claim 72 , wherein the measured optical signal along the axis may be used to determine the amount of agglutinated or unagglutinated visualization particles in the sample. 
     
     
         75 . The method of  claim 68 , wherein packed visualization particles at the bottom of the vessel are identified, and the area or perimeter of the packed visualization particles is measured. 
     
     
         76 . The method  claim 75 , wherein the method further comprises using a pattern recognition method to identify a button and, optionally, to identify a teardrop region of the packed visualization particles. 
     
     
         77 . The method of  claim 75 , wherein the area or perimeter of the packed visualization particles may be used to determine the amount of agglutinated or unagglutinated visualization particles in the sample. 
     
     
         78 . (canceled) 
     
     
         79 . The method of  claim 30 , wherein the method is performed at a location selected from a point of service location and a point of care location. 
     
     
         80 . (canceled)