Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals – Carrier is a biological cell or cell fragment
Patent
1983-06-21
1986-12-09
Marantz, Sidney
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
Carrier is a biological cell or cell fragment
422102, 436805, 436808, 436810, 436818, G01N 33555
Patent
active
046280367
DESCRIPTION:
BRIEF SUMMARY
DIAGNOSTIC TEST METHOD
The invention relates to a method for qualitatively or semi-quantitatively detecting the presence of an antigen or antibody in a test liquid, and also to a test kit which permits carrying out such a method.
The diagnosis of pathological conditions in man or animals is frequency carried out by using immunochemical reactions. In general, the presence of an antigen or antibody in a body fluid is determined immunologically by bringing into contact with corresponding antibody or antigen with the particular body fluid to be tested, usually blood serum or urine.
The presence of the antigen or antibody to be detected can then be noted by the fact that a less soluble antigen-antibody complex is formed.
Such complexes are formed rather slowly and only at relatively high concentrations of antigen and antibody. It is therefor necessary, in order to increase the sensitivity, to use carriers in order to permit the detection of this type of complex and hence also of the particular antigens and antibodies.
The coupling of antigens and antibodies to a carrier is a known technique which is described in all immunological handbooks. Extensively used carrier are, inter alia: erythrocytes, cacterial cells, bentonite, latex particles and cellulose. In the present diagnostic test met hod, use is made of erythrocytes as carriers.
The presence of, for example, an antigen in a test sample or test liquid can now be demonstrated by bringing the test sample into contact with antibodies against this antigen, which are coupled to erythrocytes. If the antigen actually is present in the test sample, an immunochemical reaction will take place between the particular antigen and the antibody which is coupled to erythrocytes, resulting in an antigen-antibody complex coupled to erythrocytes and in a coupling together (agglutination) of such erythrocyte particles; on settling out in a test tube having a round bottom, these agglutinated erythrocytes give a plain light brown pattern.
If the antigen is not present in the test liquid, no immunochemical reaction, hence also no agglutination, can take place. The antibody bonded to erythrocytes will settle out after a short time and give a characteristic ring pattern in a test tube having a round bottom.
In this test set-up, obtaining a characteristic ring pattern in the test tube therefore means that the suspected antigen is not present in the test liquid, and therefore that the test result is negative.
The antigen in question can also be detected in a different manner, namely by bringing the test sample into contact with 2 reagents:
If the antigen in question is not present in the test liquid, an immunological reaction will take place between the two added reagents, resulting in an agglutination of an erythrocyte-bonded antigen-antibody complex, which on settling out will not give a characteristic ring pattern.
If the antigen in question is present in the test liquid, this antigen will react directly with the antibody, giving a (soluble) antigen-antibody complex, while the unused erythrocyte-coupled antigen will settle out after a short time and, in a test tube with a round bottom, lead to the formation of a characteristic ring pattern.
In this test set-up, obtaining a characteristic ring pattern therefore means that the test result is positive.
Though the interpretation of the test result therefore depends on the chosen test set-up, the characteristic ring pattern of the erythrocyte-bonded antigen or antibody each time forms the basis of the test method.
Hitherto, the tests outlined above have been carried out in conventional round-bottomed test tubes. In such test tubes, the characteristic ring pattern of non-agglutinated erythrocytes manifests itself as a more or less thick ring.
Surprisingly, it has now been found that carrying out these immunochemical reactions in a test tube having a bottom in the shape of an inverted prism or truncated prism, such that the lower edge of the tube forms a strip having a length which is at least equal to one-fourth of the distance between
REFERENCES:
patent: 4148607 (1979-04-01), Bernoco
patent: 4290997 (1981-09-01), Suovaniemi
patent: 4297104 (1981-10-01), Matte
patent: 4303616 (1981-12-01), Kano
patent: 4427634 (1984-01-01), Truglio
patent: 4466740 (1984-08-01), Kano
Patent Abstracts of Japan, vol. 4, No. 95, Jul. 9, 1980, p. 19P18.
Scheepens Antonius H. J. M.
Theunissen Hendrikus J. J.
van Hell Hans
Akzo N.V.
Marantz Sidney
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