Multi-immunoassay diagnostic system for antigens or antibodies or both

• US 5,447,837 A
• Filed: 02/06/1989
• Issued: 09/05/1995
• Est. Priority Date: 08/05/1987
• Status: Expired due to Fees

• Current Assignee: Calypte Incorporated
• Original Assignee: Calypte Incorporated
• Inventors: Urnovitz, Howard B.
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US07/307,361
• Time in Patent Office: 2,402 Days
• Field of Search: 435/7, 435/805, 435/810, 435/7.9, 435/7.92, 435/7.95, 435/973, 435/974, 435/5, 435/962, 436/501, 436/514, 436/518, 436/523, 436/524, 436/528, 436/807, 436/531, 436/810, 436/, 436/825, 436/820, 436/513, 422/55
• US Class Current: 435/5
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 435/974 Aids related test
• CPC Class Codes: Y10S 436/825 Pretreatment for removal of interfering factors from sample

Abstract

The present invention provides a test strip for detecting, in a sample from a human subject, the presence of an antigenic substance which comprises a solid support, an antibody directed against the antigenic substance bound to a first discrete area on the solid support, an anti-human antibody bound to a second discrete area on the solid support as a positive control, and an antibody directed against an antigen which does not naturally occur in human subjects bound to a third discrete area on the solid support as a negative control. The present invention also provides a test strip for detecting, in a sample from a human subject, the presence of an antibody which comprises a solid support, an antigenic substance bound to a first discrete area on the solid support, an anti-human antibody bound to a second discrete area on the solid support as a positive control, and a negative control bound to a third discrete area on the solid support. The invention further provides a test strip for detecting the presence of an antibody and antigenic substance, which may additionally comprise an antibody-based reagent directed against the antigenic substance in an immune complex and native human antibody thereto. The invention also provides a method for detecting in a sample from a human subject the presence of an antigenic substance or antibody or both using the aforementioned test strips.

Claims

1. A kit for detecting the presence of a target human antibody to human immunodeficiency virus (HIV) in a urine sample comprising

a) a treatment buffer comprising non-immune sera and about 0.01% to 0.5% (w/v) of a plurality of solid phase particles from about 0.5 microns to about 10 microns in diameter, the plurality of solid phase particles comprising equal volumes of three particle types, each particle type coated with goat, bovine, or horse immunoglobulin antibodies, the non-immune sera comprising 3% bovine serum, 3% goat serum, and 3% horse serum;

b) a labelled reagent comprising an enzyme label conjugated to an anti-human immunoglobulin antibody;

c) a substrate specific for the enzyme label; and

d) a reagent HIV antigen.

2. A buffer comprising non-immune sera and about 0.01% to 0.5% (w/v) of a plurality of solid phase particles from about 0.5 microns to about 10 microns in diameter, the plurality of solid phase particles comprising equal volumes of three particle types, each particle type coated with goat, bovine, or horse immunoglobulin antibodies, the non-immune sera comprising 3% bovine serum, 3% goat serum, and 3% horse serum.

3. A method for detecting the presence of a target human antibody to HIV in a urine sample, said method comprising:

a) adding a treatment buffer to the sample, which buffer comprises non-immune sera and 0.01% to 0.5% (w/v) of a plurality of solid phase particles from about 0.5 microns to about 10 microns in diameter, the plurality of solid phase particles comprising equal volumes of three particle types, each particle type coated with goat, bovine, or horse immunoglobulin antibodies, the non-immune sera comprising 3% bovine serum, 3% goat serum, and 3% horse serum; and

b) contacting the sample with a reagent HIV antigen to form an antigen-antibody complex containing the target human antibody and the HIV antigen;

c) contacting the antigen-antibody complex with an enzyme labeled anti-human immunoglobulin antibody that specifically binds to the target human antibody; and

d) detecting the presence of the bound label as an indication of the presence of any target antibody in the urine sample.

4. The method of claim 3 wherein the HIV antigen is gp160, gp120 or gp41 glycoprotein, or p24 protein.

5. A method of detecting an HIV antibody in a saliva, urine, or whole or fractionated blood sample from a human subject, said method comprising:

a) contacting the sample with a recombinant HIV glycoprotein under conditions such that the glycoprotein specifically binds to any HIV antibody present in the sample to form a complex;

b) contacting the complex with an enzyme labeled anti-human immunoglobulin antibody which specifically binds and labels the complex to form a labeled complex; and

c) detecting the presence of enzyme labeled complex and thereby the presence of any HIV antibody in the sample wherein a treatment buffer is added to the sample before, or simultaneous with, contacting the sample with the glycoprotein, said treatment buffer comprising non-immune sera and about 0.01% to 0.5% (w/v) of a plurality of solid phase particles from about 0.5 microns to about 10 microns in diameter, the plurality of solid phase particles comprising equal volumes of three particle types, each particle type coated with goat, bovine, or horse immunoglobulin antibodies, the non-immune sent comprising 3% bovine serum, 3% goat serum, and 3% horse serum.

6. A method for detecting in a sample from a human subject the presence of a target human antibody to HIV which specifically binds an HIV vital antigen, said method comprising:

a) adding to the sample a treatment buffer comprising non-immune sera and about 0.01% to 0.5% (w/v) of a plurality of solid phase particles from about 0.5 microns to about 10 microns in diameter, the plurality of solid phase particles comprising equal volumes of three particle types, each particle type coated with goat, bovine, or horse immunoglobulin antibodies, the non-immune sent comprising 3% bovine serum, 3% goat serum, and 3% horse serum;

b) using a test strip comprising

i) a solid support;

ii) said HIV viral antigen bound to a first discrete area on the solid support;

iii) a non-target human antibody bound to a second discrete area on the solid support as a positive control; and

iv) a negative control which will not specifically bind target human antibody or antihuman antibody bound to a third discrete area on the solid support;

c) contacting the treated sample with the test strip under conditions such that the HIV viral antigen bound to the test strip specifically binds with any target human antibody present in the treated sample;

d) washing the test strip to remove unbound treated sample;

e) contacting the resulting test strip with enzyme labeled antihuman antibodies which specifically bind to any target human antibodies bound to, or on, the test strip;

f) detecting the presence of enzyme labeled antibodies and thereby the presence of target human antibodies in the sample; and

g) verifying the correctness of the detection by determining that the positive control is enzyme labeled and the negative control is not enzyme labeled.

7. The method of claim 6 wherein the sample comprises urine, saliva, or whole or fractionated blood.

8. A method for detecting in a sample from a human subject the presence of a target human antibody to HIV which specifically bins an HIV viral antigen, said method comprising:

a) adding to the sample a treatment buffer comprising non-immune sera and about 0.01% to 0.5 (w/v) of a plurality of solid phase particles from about 0.5 microns to about 10 microns in diameter, the plurality of solid phase particles comprising equal volumes of three particle types each particle type coated with goat, bovine, or horse immunoglobulin antibodies, the non-immune sen comprising 3% bovine serum, 3% goat serum, and 3% horse serum;

b) using a test strip comprising;

i) wells used as a solid support;

ii) said HIV viral antigen bound to discrete areas on the solid support;

c) contacting the treated sample with the test strip under conditions such that the HIV viral antigen bound to the test strip specifically binds with any target human antibody present in the treated sample;

d) washing the test strip to remove unbound treated sample;

e) contacting the resulting test strip with enzyme labeled antihuman antibodies which specifically bind to any target human antibodies bound to, or on, the test strip; and

f) detecting the presence of any bound enzyme labeled antibodies using a substrate for the enzyme and thereby the presence of the target human antibodies in the treated sample.

9. The method of claim 8, wherein the HIV viral antigen is gp160, gp120 or gp41 glycoprotein, or p24 protein.

10. The method of claim 8 wherein the HIV viral antigen is recombinant gp 160.