Processes for the detection of non-A, non-B hepatitis and a kit for use in the same

Claim 2

A process for assaying a tissue specimen for non-A, non-B hepatitis antigen, which process comprises: (a) providing a tissue specimen to be assayed; (b) contacting the tissue specimen with a non-A, non-B hepatitis antibody; (c) contacting the resultant tissue specimen with a biotin-avidin system; and (d) determining the presence of non-A, non-B hepatitis antigen present in the tissue sample.

Claim 3

A process according to Claim 1, wherein the tissue specimen is a piece of liver tissue.

Claim 4

A process according to Claim 2, wherein the tissue specimen is fixed and embedded.

Claim 5

A process according to Claim 2, whrein the tissue specimen is unfixed, frozen tissue.

Claim 6

A process according to any preceding claim, wherein the tissue specimen is pretreated or otherwise contacted with a blocking agent to block nonspecific binding sites, the blocking agent preferably being chosen from: normal IgG, normal IgM, bovine serum albumin, and normal animal serum.

Claim 7

A process according to any preceding claim, wherein the non-A, non-B hepatitis antibody is IgG or IgM.

Claim 8

A process according to any preceding claim, wherein the tissue specimen is contacted with the non-A, non-B hepatitis antibody at a temperature of up to 45°C, preferably from 20°C to 37°C, for a time of up to 24 hours, preferably from 2 to 24 hours.

Claim 9

A process according to any preceding claim, wherein the non-A, non-B hepatitis antibody of step (b) is biotinylated prior to its contacting with the tissue specimen.

Claim 10

A process according to any preceding claim, wherein the resultant tissue specimen of step (c) is contacted with a biotin labelled antibody against the non-A, non-B hepatitis antibody which biotin labelled antibody is derived from a host distinct from the host sued to produce the non-A, non-B hepatitis antibody.

Claim 11

A process according to any one of Claims 1 to 7, wherein the resultant tissue specimen of step (c) is first contacted with a biotin labelled antibody and then contacted with readable labelled avidin-biotin system.

Claim 12

A process for assaying for non-A, non-B hepatitis antibody present in a biological sample, which process comprises: (a) contacting a tissue specimen containing non-A, non-B hepatitis antigen with the biological specimen to permit reaction of the specimen non-A, non-B hepatitis antibodies with the non-A, non-B hepatitis antigen; (b) incubating the reactants for a time of up to 24 hours at a temperature of up to 45°C; (c) separating the tissue specimen from the non-reactive phase; (d) contacting the resultant tissue specimen with a biotin-avidin system; and (e) determining the presence of non-A, non-B hepatitis antibody in the sample.

Claim 13

A process for assaying for non-A, non-B hepatitis antibody present in a biological sample, which process comprises: (a) contacting a tissue specimen containing non-A, non-B hepatitis antigen with the biological specimen to permit reaction of the specimen non-A, non-B hepatitis antibodies with the non-A, non-B hepatitis antigen; (b) incubating the reactants for a time of up to 24 hours at a temperature of up to 45°C; (c) separating the tissue specimen from the non-reactive phase; (d) contacting the resultant tissue specimen with a biotin-avidin system; and (e) determining the presence of non-A, non-B hepatitis antibody in the sample.

Claim 14

A process according to Claim 11, wherein a positive result indicates the presence of non-A, non-B hepatitis antibody in the sample.

Claim 15

A process for assaying for non-A, non-B hepatitis anitgen present in a biological sample, which process comprises: (a) reacting antigen present in the sample with non-A, non-B hepatitis antibody for a time of up to 10 days at a temperature of up to 45°C; (b) separating reactive phase from non-reactive phase by precipitating immune-complexed antibody - antigen and recovering the liquid supernatant; (c) contacting a tissue specimen containing non-A, non-B hepatitis antigen with the liquid supernatant to-permit reaction of any non-precipitated non-A, non-B hepatitis antibodies with the non-A, non-B hepatitis antigen present in the tissue specimen; (d) incubating the reactants for a period of up to 24 hours at a temperature of up to 45°C; (e) contacting the resultant tissue specimen with a biotin-avidin system; and (f) determining the presence of non-A, non-B hepatitis antigen in the sample.

Claim 16

A process for assaying for non-A, non-B hepatitis anitgen present in a biological sample, which process comprises: (a) reacting antigen present in the sample with non-A, non-B hepatitis antibody for a time of up to 10 days at a temperature of up to 45°C; (b) separating reactive phase from non-reactive phase by precipitating immune-complexed antibody - antigen and recovering the liquid supernatant; (c) contacting a tissue specimen containing non-A, non-B hepatitis antigen with the liquid supernatant to-permit reaction of any non-precipitated non-A, non-B hepatitis antibodies with the non-A, non-B hepatitis antigen present in the tissue specimen; (d) incubating the reactants for a period of up to 24 hours at a temperature of up to 45°C; (e) contacting the resultant tissue specimen with a biotin-avidin system; and (f) determining the presence of non-A, non-B hepatitis antigen in the sample.

Claim 17

A process according to Claim 13, wherein a negative result indicates the presence of non-A, non-B hepatitis antigen in the sample.

Claim 18

A process according to Claim 13 or 14, wherein the reaction of anitgen with antibody in step (a) is performed in two steps, the first step being at a temperature of from 20 to 45°C for from 1 to 5 hours and a second step at a temperature lower than the temperature of the first step and for a time longer than that of the first step.

Claim 19

A tissue specimen which is fixed and embedded and which contains non-A, non-B hepatitis antigen.

Claim 20

A tissue specimen which is fixed and embedded and which contains non-A, non-B hepatitis antigen.

Claim 21

A diagnostic kit which comprises: (a) a tissue specimen which is fixed and embedded and which specimen contains non-A, non-B hepatitis antigen; (b-) a quantity of non-A, non-B hepatitis antibody; (c) a quantitiy of a biotin-avidin system; and (d) a substrate for the determination of the enzyme activity of the enzyme.

Claim 22

A diagnostic kit which comprises: (a) a tissue specimen which is fixed and embedded and which specimen contains non-A, non-B hepatitis antigen; (b-) a quantity of non-A, non-B hepatitis antibody; (c) a quantitiy of a biotin-avidin system; and (d) a substrate for the determination of the enzyme activity of the enzyme.

Claim 23

A diagnostic kit according to Claim 17, wherein the biotin-avidin system comprises a biotin labelled antibody and a readable labelled avidin-biotin system.