Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals – Carrier is a biological cell or cell fragment
Patent
1995-06-26
1998-01-13
Housel, James C.
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
Carrier is a biological cell or cell fragment
436 66, 435 14, G01N 33555, G01N 3324
Patent
active
057078785
DESCRIPTION:
BRIEF SUMMARY
TECHNOLOGICAL FIELD
The present invention relates to a method for specifically detecting blood component derived from human and, more particularly, to a method for detecting a blood component capable of detecting minute bleeding in digestive tract for the purposes of screening, for example, the screening of colorectal cancer and the like, and a kit using this detection method.
BACKGROUND ART
Detection of blood component of human is required in a variety of fields. The necessity is especially important in the clinical field. Bleeding, for example bleeding in digestive tract, occurs in a variety of diseases. Particularly, it is well known that bleeding from digestive tracts is an important early stage symptom in malignant tumors of digestive tracts.
Detection of blood components in feces, especially hemoglobin, is a major method currently applied to the detection of digestive tract bleeding for diagnosis of diseases such as malignant tumors of digestive tracts. Reversed passive hemagglutination, latex agglutination (fixation), gold colloid agglutination, enzyme immunoassay, radio immunoassay, and the like are included in the methods for detecting the hemoglobin in stools. Their principle is preparing an anti-human hemoglobin antibody, causing an antigen-antibody reaction with hemoglobin, and measuring or detecting the resultant by various methods.
In the conventional immunological method for detecting hemoglobin using the anti-human hemoglobin antibody, however, the hemoglobin to be detected is usually supplied to the test together with feces which act as an inactivating factor. The antigenic determinant of hemoglobin may be inactivated depending on the temperature over time, resulting in a drawback of conspicuous deterioration of the detection sensitivity.
Hemoglobin may be denatured or decomposed, particularly under high temperature conditions in summertime, therefore, hemoglobin positive feces are often reduced to negative in summer {Takefumi FUJIYOSHI and Mariko 23(15), 6097-6101 (1989), "Basic studies on immune stool occult blood reagent"}. The inactivation of hemoglobin is significant particularly when its concentration is low. It is difficult to perform hemoglobin detection which is meaningful for diagnosis in such low concentration levels.
Development of a method for properly measuring blood component in samples of feces or the like without being affected by temperatures, storage time and the like has been required. The object of the present invention is to provide such a method.
DISCLOSURE OF THE INVENTION
The present inventors have conducted extensive studies in order to detect blood or blood component in fecal samples or the like using a blood component other than hemoglobin.
As a result, the inventors have found that human erythrocyte membrane band 3 glycoprotein (hereinafter referred to as band 3) can be properly detected, because it is much mores table than hemoglobin even in samples of feces or the like, and forms a complex by binding with a certain hemagglutinin specific for human erythrocytes. This finding has led to the completion of the present invention.
Accordingly, the present invention provides a method for detecting a blood component in a sample comprising reacting band 3 in the sample and a hemagglutinin produced by a microorganism belonging to the genus Conidiobolus and measuring the complex produced by the reaction, and a kit used for the detection.
BEST MODE FOR CARRYING OUT THE INVENTION
Band 3 which is detected as a blood component in the method of the present invention is a component which has already been reported as an erythrocyte marker (Beppu, M. et al., J. Biol. Chem., 256(6), 3226-3233 (1990), "Binding of anti-band 3 autoantibody to oxidatively damaged erythrocytes", etc.) and been studied as useful for monitoring erythrocytes.
The hemagglutinin produced by a microorganism belonging to the genus Conidiobolus (hereinafter abbreviated as CA) is a human erythrocyte-specific hemagglutinin produced by a microorganism belonging to the genus Conidiobolus such as Conidi
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Kotsugai Takeshi
Narita Shigeo
Narita Tadashi
Tomiyama Tetsuo
Housel James C.
Portner Ginny Allen
SS Pharmaceutical Co., Ltd.
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