FXIII detection for verifying serum sample and sample size and for detecting dilution

Abstract

Analyses of serum samples for the presence and amount of either of the two subunits of human Factor XIII protein are used as a means of eliminating a significant source of error that arises in the testing of serum and plasma. For serum samples, a negative result of an analysis for the presence of subunit a is a means of verifying that a sample is indeed serum, while a negative or positive result for subunit a serves to distinguish serum (negative) from plasma (positive). A positive result for the presence of subunit b is a means of verifying that the sample is either serum or plasma and not any other biological fluid. A quantitative analysis of subunit b is a means of verifying that the sample is of the intended volume rather than having been reduced in volume due to improper sampling. A quantitative analysis of subunit b is also a means of verifying the dilution of a sample of either serum or plasma.

What is claimed is:  
     
         1 . A method for verifying that an analytical test sample to be analyzed for the presence or absence of a biological condition is a sample of human serum and does not contain human plasma, said method comprising analyzing said sample for the presence therein of the a subunit of human Factor XIII protein, any a subunit thus detected being an indication of the presence of human plasma in said sample.  
     
     
         2 . A method in accordance with  claim 1  in which the presence of said a subunit is determined by immunoassay.  
     
     
         3 . A method in accordance with  claim 2  in which said immunoassay comprises contacting said sample with a capture antibody that has specific binding affinity for said a subunit and substantially no binding affinity toward the b subunit of human Factor XIII protein, and detecting said a subunit thus captured.  
     
     
         4 . A method in accordance with  claim 2  in which said immunoassay comprises (i) contacting said sample with a solid phase to which is bound a capture antibody that has specific binding affinity for said a subunit and substantially no binding affinity toward the b subunit of said human Factor XIII protein, and (ii) determining whether any binding of said a subunit to said solid phase through said capture antibody has occurred.  
     
     
         5 . A method in accordance with  claim 4  in which said solid phase is a population of particles, and (ii) comprises detecting said particles by flow cytometry.  
     
     
         6 . A method in accordance with  claim 2  in which said immunoassay comprises contacting said sample with a capture antibody that has binding affinity for said a subunit, and detecting said a subunit thus captured with a detector antibody that has specific binding affinity for said a subunit and substantially no binding affinity toward the b subunit of human Factor XIII protein.  
     
     
         7 . A method in accordance with  claim 6  in which said immunoassay comprises (i) contacting said sample with a solid phase to which is bound a capture antibody that has binding affinity for said a subunit, and (ii) determining whether any binding of said a subunit to said solid phase through said capture antibody has occurred by detecting any a subunit thus bound with a detector antibody that has specific binding affinity for said a subunit and substantially no binding affinity toward the b subunit of said human Factor XIII protein.  
     
     
         8 . A method in accordance with  claim 6  in which said solid phase is a population of particles, and (ii) comprises detecting said particles by flow cytometry.  
     
     
         9 . A method for verifying that an analytical test sample to be analyzed for the presence or absence of a biological condition is a sample of either serum or plasma and is not a sample of either cerebrospinal fluid, urine or non-human serum, said method comprising analyzing said sample for the presence of the b subunit of human Factor XIII protein in said sample, the presence of said b subunit verifying that said sample is a sample of human serum or plasma.  
     
     
         10 . A method in accordance with  claim 9  in which the presence of said b subunit is determined by immunoassay.  
     
     
         11 . A method in accordance with  claim 10  in which said immunoassay comprises contacting said sample with a capture antibody that has specific binding affinity for said b subunit, and detecting said b subunit thus captured.  
     
     
         12 . A method in accordance with  claim 10  in which said immunoassay comprises (i) contacting said sample with a solid phase to which is bound a capture antibody that has specific binding affinity for said b subunit, and (ii) determining whether any binding of said b subunit to said solid phase through said capture antibody has occurred.  
     
     
         13 . A method in accordance with  claim 12  in which said solid phase is a population of particles, and (ii) comprises detecting said particles by flow cytometry.  
     
     
         14 . A method for determining the degree of dilution of a sample of either serum or plasma, said method comprising analyzing said sample for the amount of the b subunit of human Factor XIII protein in said sample, and comparing said amount with the amount of b subunit in a sample of human serum or plasma of known degree of dilution.  
     
     
         15 . A method in accordance with  claim 14  in which the amount of said b subunit in said sample is determined by immunoassay.  
     
     
         16 . A method in accordance with  claim 15  in which said immunoassay comprises contacting said sample with a capture antibody that has specific binding affinity for said b subunit, and detecting said b subunit thus captured.  
     
     
         17 . A method in accordance with  claim 15  in which said immunoassay comprises (a) contacting said sample with a solid phase to which is bound a capture antibody that has specific binding affinity for said b subunit, and (b) determining whether any binding of said b subunit to said solid phase through said capture antibody has occurred.  
     
     
         18 . A method in accordance with  claim 17  in which said solid phase is a population of particles, and (b) comprises detecting said particles by flow cytometry.  
     
     
         19 . A method for verifying that a sample of either serum or plasma is of a selected volume, said method comprising analyzing said sample for the amount of the b subunit of human Factor XIII protein in said sample, and comparing said amount thus detected with the amount of b subunit in a sample of human serum or plasma known to be of said selected volume.  
     
     
         20 . A method in accordance with  claim 19  in which the amount of said b subunit is determined by immunoassay.  
     
     
         21 . A method in accordance with  claim 20  in which said immunoassay comprises contacting said sample with a capture antibody that has specific binding affinity for said b subunit, and detecting said b subunit thus captured.  
     
     
         22 . A method in accordance with  claim 20  in which said immunoassay comprises (a) contacting said sample with a solid phase to which is bound a capture antibody that has specific binding affinity for said b subunit, and (b) determining whether any binding of said b subunit to said solid phase through said capture antibody has occurred.  
     
     
         23 . A method in accordance with  claim 22  in which said solid phase is a population of particles, and (b) comprises detecting said particles by flow cytometry.