EP0200507A1 — Improved enzyme immunoassay and a kit therefor

Claim 1

Independent Claim 81 words

The use, in an enzyme immunoassay, of a detecting support carrying thereon enzyme detecting reagent fixed at a first set of localised predetermined positions in detecting enzyme labels immobilized at a corresponding and second set of fixed localised predetermined positions carried on an immunological reaction support, the immobilization of which labels is the result of an immunoassay reaction, said detecting support being superimposible over said reaction support with the respective sets of positions in contact to permit detection of enzyme labels.

Claim 2

Dependent Claim 81 words

The use, in an enzyme immunoassay, of a detecting support carrying thereon enzyme detecting reagent fixed at a first set of localised predetermined positions in detecting enzyme labels immobilized at a corresponding and second set of fixed localised predetermined positions carried on an immunological reaction support, the immobilization of which labels is the result of an immunoassay reaction, said detecting support being superimposible over said reaction support with the respective sets of positions in contact to permit detection of enzyme labels.

Claim 3

Independent Claim 167 words

A method for assaying an immunologically active substance (B) in a test sample which comprises; (a) preparing an immuno-active membrane of a hydrophobic polymer sheet having a number of hydrophilic spots thereon, onto which a prefixed amount of an immuno-active substance (A) or (B) has been immobilized, (b) preparing a detecting-membrane having a number of spots which contain a detectable reagent which can be detected by the catalytic effect of an enzyme at positions corresponding to those of the immuno-active membrane, (c) making contact the immuno-active membrane with the test sample together with an enzyme-labelled immuno-active substance (A') or (B'), simultaneously or successively, whereby reacting the substance (A), substance (B) and enzyme-labelled substance (A') or (B') competitively or non-competitively, (d) adding a substrate to each spot of the immuno-active membrane, (e) superimposing the detecting membrane on the immuno-active membrane so that the spots in both membranes correspond to each other, (f) detecting the detectable reagent and thereby determining the amount of substance (B) in the test sample.

Claim 4

Independent Claim 167 words

A method for assaying an immunologically active substance (B) in a test sample which comprises; (a) preparing an immuno-active membrane of a hydrophobic polymer sheet having a number of hydrophilic spots thereon, onto which a prefixed amount of an immuno-active substance (A) or (B) has been immobilized, (b) preparing a detecting-membrane having a number of spots which contain a detectable reagent which can be detected by the catalytic effect of an enzyme at positions corresponding to those of the immuno-active membrane, (c) making contact the immuno-active membrane with the test sample together with an enzyme-labelled immuno-active substance (A') or (B'), simultaneously or successively, whereby reacting the substance (A), substance (B) and enzyme-labelled substance (A') or (B') competitively or non-competitively, (d) adding a substrate to each spot of the immuno-active membrane, (e) superimposing the detecting membrane on the immuno-active membrane so that the spots in both membranes correspond to each other, (f) detecting the detectable reagent and thereby determining the amount of substance (B) in the test sample.

Claim 5

Dependent Claim 33 words

A method as claimed in claim 2 wherein the hydrophilic spots are formed on the hydrophobic polymer sheet by means of a low temperature plamsa-treatment method using oxygen, oxygen-containing gas or water vapour.

Claim 6

Dependent Claim 17 words

A method as claimed in claim 2 or claim 3 wherein the hydrophobic polymer sheet is porous.

Claim 7

Independent Claim 23 words

A method as claimed in any one of claims 2 to 4 wherein the hydrophobic polymer sheet is a polyolefin sheet, e.g. polypropylene.

Claim 8

Independent Claim 21 words

A method as claimed in any one of claims 2 to 5 wherein detection of the reaction is carried out colorimetrically.

Claim 9

Dependent Claim 26 words

A method as claimed in claim 6 wherein a colouring-membrane is prepared by immobilizing a pigment at the regions corresponding to those of the hydrophilic spots.

Claim 10

Independent Claim 31 words

A method as claimed in any one of claims 2 to 7 wherein the assay is conducted as a sandwich assay as a competitive assay or as a competitive hindrance assay.

Claim 11

Independent Claim 25 words

A method as claimed in any one of claims 2 to 8 wherein the hydrophilic spots are circular and of 0.5 to 15 mm diameter.

Claim 12

Dependent Claim 23 words

An immunoassay kit which comprises an immuno-active membrane, a supporting board and a flame board in a suitable container and a detecting membrane.

Claim 13

Dependent Claim 23 words

An immunoassay kit which comprises an immuno-active membrane, a supporting board and a flame board in a suitable container and a detecting membrane.

Claim 14

Dependent Claim 36 words

The use of an immuno-assay kit as claimed in claim 10 in the determination of an immunologically active substance in a biological sample e.g in diagnosis other than when practised on the human or animal body.

Improved enzyme immunoassay and a kit therefor

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