Immunologic test method and immunologic test kit

Details Immunologic test method and immunologic test kit Immunologic test method and immunologic test kit

2000-02-29

2004-06-22

Chin, Christopher L. (Department: 1641)

Chemistry: analytical and immunological testing

Involving an insoluble carrier for immobilizing immunochemicals

C436S523000, C436S525000, C436S528000, C436S535000, C436S808000, C435S007100, C435S007940, C435S007920, C435S007500, C435S004000

Reexamination Certificate

active

06753190

ABSTRACT:

TECHNICAL FIELD
The present invention relates to an immunological detection method capable of performing detection of an analyte in a test sample and a kit used therefor. More particularly, the present invention relates to immunochromatography comprising the step of amplifying a detection signal.
BACKGROUND ART
In an immunological analysis of a biological sample, or the like, a method for carrying out its detection quickly and simply includes immunochromatography. This method generally comprises the steps as described below. Concretely, when a mixture comprising a liquid of a test sample and a labeled antibody capable of specifically binding to an analyte is absorbed and developed from one end of a test strip comprising a water-absorbent substrate having a capture region immobilized with an antibody capable of specifically binding to the analyte, a labeled antibody-analyte complex formed in the mixture is bound to the immobilized antibody, thereby capturing the complex on the capture region. Therefore, the analyte in the liquid of a test sample can be determined by assaying the labeled antibody bound to the capture region.
In addition, as a method for obtaining a detection signal by the immunochromatography mentioned above with a higher sensitivity, a method using two kinds of labeled antibodies is disclosed in Japanese Patent Laid-Open No. Hei 10-062419. In other words, the construction is such that a first labeled antibody and a second labeled antibody are respectively positioned (absorbed) as reagent regions between a capture region (immobilized with another antibody capable of specifically binding to the analyte) and a site at which a test sample is absorbed in the test strip, wherein the first labeled antibody is obtained by labeling an antibody capable of specifically binding to an analyte, and the second labeled antibody is obtained by labeling a secondary antibody capable of specifically binding to the antibody. The analyte in the sample forms a complex with the first labeled antibody, and thereafter the second labeled antibody is further bound to the first labeled antibody, thereby forming a complex of [analyte-first labeled antibody-second labeled antibody]. The immunological complex is captured by an antibody immobilized on the capture region. Therefore, a signal amplified by the second labeled antibody is detected on the capture region.
However, at present, a sufficient detection sensitivity has not yet been obtained even by the signal-amplifying immunochromatography described above. Further, when a test sample is feces, urine, blood, or the like, there is necessitated additional procedures such as a step of suspending a sample in an appropriate buffer as a pretreatment and/or partial purification process comprising separating and removing heterogeneous substances in the test sample, so that it has a defect of lack of quickness.
Accordingly, an object of the present invention is to provide an immunological detection method relating to immunochromatography, the method capable of detecting an analyte more quickly and with high sensitivity, and a kit used therefor.
DISCLOSURE OF INVENTION
The present invention provides the following immunological detection methods and kits used therefor:
[1] an immunological detection method for detecting an analyte by using a water-absorbent substrate in which a capture region immobilized with a first immunochemical component capable of specifically binding to the analyte is positioned in a given region on a surface thereof, wherein the immunological detection method is characterized by the use of:
(1) a solution comprising a labeled immunochemical component (first labeled immunochemical component) comprising a second immunochemical component capable of specifically binding to the analyte and a labeling substance, wherein the labeling substance is bound to the second immunochemical component; and
(2) a solution comprising a labeled immunochemical component (second labeled immunochemical component) comprising a third immunochemical component capable of specifically binding to the second immunochemical component and a labeling substance, wherein the labeling substance is bound to the third immunochemical component;
[2] an immunological detection method comprising forming on a capture region an immunological complex in which an analyte in a test sample is sandwiched with a first immunochemical component capable of specifically binding to the analyte, and a labeled component (first labeled component), the first labeled component comprising a second immunochemical component capable of specifically binding to the analyte, a third immunochemical component incapable of binding to the analyte and a labeling substance, wherein the labeling substance is bound to the second and third immunochemical components, wherein the first immunochemical component is immobilized on the capture region positioned in a given region on a surface of a water-absorbent substrate; and determining a signal of the labeling substance on the capture region, characterized in that the method comprises forming an immunological complex in which a labeled component (second labeled component) is bound to the third immunochemical component present in a sandwiched immunological complex via a mediating substance, the second labeled component comprising a fourth immunochemical component capable of specifically binding to the third immunochemical component via the mediating substance, and a labeling substance, wherein the labeling substance is bound to the fourth immunochemical component, thereby amplifying the signal of the labeling substance;
[3] a sandwiched-type immunological detection method wherein at a capture region immobilized with a first immunochemical component capable of binding to an analyte, the analyte is sandwiched by the first immunochemical component and a labeled component comprising a second immunochemical component capable of binding to the analyte and a labeling substance, wherein the labeling substance is bound to the second immunochemical component, characterized in that the immunological detection method comprises forming a complex via binding between a biotin and an avidin, and detecting the analyte;
[4] a kit for immunological detection characterized in that the kit comprises a water-absorbent substrate in which a capture region immobilized with a first immunochemical component capable of specifically binding to an analyte is positioned in a given region on a surface thereof; a labeled immunochemical component (first labeled immunochemical component), the first labeled immunochemical component comprising a second immunochemical component capable of specifically binding to the analyte, and a labeling substance, wherein the labeling substance is bound to the second immunochemical component; and a labeled immunochemical component (second labeled immunochemical component), the second labeled immunochemical component comprising a third immunochemical component capable of specifically binding to the second immunochemical component, and a labeling substance, wherein the labeling substance is bound to the third immunochemical component;
[5] a kit for immunological detection comprising a water-absorbent substrate in which a capture region immobilized with a first immunochemical component capable of specifically binding to an analyte is positioned in a given region on a surface thereof; a labeled component (first labeled component), the first labeled immunochemical component comprising a second immunochemical component capable of specifically binding to the analyte, a third immunochemical component incapable of binding to the analyte, and a labeling substance, wherein the labeling substance is bound to the second and third immunochemical components; a labeled component (second labeled component), the second labeled component comprising a fourth immunochemical component capable of specifically binding to the third immunochemical component via a mediating substance, and a labeling substance, wherein the

Affiliated with

Also associated with

Rating

Immunologic test method and immunologic test kit does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Immunologic test method and immunologic test kit, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Immunologic test method and immunologic test kit will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3365412