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US4764459A
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Enzyme-linked immunosorbent assay (ELISA) for determining anti-bodies against herpes simplex virus (HSV) types 1 and 2 in human sera
Assignee: United States, HEALTH AND HUMAN SERVICES THE, Secretary of
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Filing: 1984-12-28
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Granted: 1988-08-16
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Expires: 2004-12-28
Abstract
Abstract A method and test kit for the serological diagnosis of human infection by herpes sipmlex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) using immunoaffinity purified virus-coded glycoproteins as target antigens. A preferred embodiment of the method employs a variation of the enzyme-linked immunosorbent assay (ELISA) whereby monoclonal antibodies are used to purify target antigens, and test sera are absorbed with virus-infected cell extracts to remove intertypic cross-reacting antibodies.
Inventors
- 👤 Berge Hampar
- 👤 Stephen D. Showalter
- 👤 Martin Zweig
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 20:56
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; G01N33/56983; G01N33/56994; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S436/00; Y10S436/807; Y10S436/809; Y10S436/825
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
87 words
An immunoassay method for determining the presence of antiboides directed against HSV-1 in a biological fluid comprising: (a) absorbing said biological fluid with an extract from HSV-2 infected cell culture under conditions wherein intertypic cross-reactive antibodies are removed; (b) reacting the biological fluid of step "a" with a target antigen, wherein said target antigen is a purified glycoprotein encoded by HSV-1, and wherein said target antigen specifically binds to antibodies contained in said biological fluid; and (c) measuring the amount of antibody bound to said target antigen.
Claim 2
Dependent Claim
87 words
An immunoassay method for determining the presence of antiboides directed against HSV-1 in a biological fluid comprising: (a) absorbing said biological fluid with an extract from HSV-2 infected cell culture under conditions wherein intertypic cross-reactive antibodies are removed; (b) reacting the biological fluid of step "a" with a target antigen, wherein said target antigen is a purified glycoprotein encoded by HSV-1, and wherein said target antigen specifically binds to antibodies contained in said biological fluid; and (c) measuring the amount of antibody bound to said target antigen.
Claim 3
Dependent Claim
22 words
A method of claim 1 wherein said target antigen is purified HSV-1 gC or gD or a mixture of gC and gD.
Claim 4
Dependent Claim
87 words
An immunoassay method for determining the presence of antibodies directed against HSV-2 in a biological fluid comprising: (a) absorbing said biolgical fluid with an extract from HSV-1 infected cell culture under conditions wherein intertypic cross-reactive antibodies are removed; (b) reacting the biological fluid of step "a" with a target antigen, wherein said target antigen is a purified glycoprotein encoded by HSV-1, and wherein said target antigen specifically binds to antibodies contained in said biological fluid; and (c) measuring the amount of antibody bound to said target antigen.
Claim 5
Dependent Claim
87 words
An immunoassay method for determining the presence of antibodies directed against HSV-2 in a biological fluid comprising: (a) absorbing said biolgical fluid with an extract from HSV-1 infected cell culture under conditions wherein intertypic cross-reactive antibodies are removed; (b) reacting the biological fluid of step "a" with a target antigen, wherein said target antigen is a purified glycoprotein encoded by HSV-1, and wherein said target antigen specifically binds to antibodies contained in said biological fluid; and (c) measuring the amount of antibody bound to said target antigen.
Claim 6
Dependent Claim
22 words
A method of claim 2 wherein said target antigen is purified HSV-2 gD or gF or a mixture of gD and gF.
Claim 7
Dependent Claim
134 words
An enzyme-linked immunosorbent assay (ELISA) for diagnosing human infection caused by HSV-1 which comprises: (a) immobilizing a target antigen which is a mixture of monoclonal antibody immunoaffinity purifed HSV-1 gC and gD as HSV-1 antigen; (b) absorbing human test serum with HSV-2-infected cell culture extract; (c) reacting said target antigen with the human test serum of step "b" to form a first antigen-antibody complex; (d) reacting said first complex with an enzyme-linked anti-human immunoglobulin to form a second complex; (e) reacting said second complex with a color-producing solution specific for the enzyme in step "c"; (f) repeating steps b-e with a second portion of test serum and extract from non-infected cell culture; and (g) measuring the ratio of absorbance generated by the human serum of step "b" to the human serum of step "f".
Claim 8
Dependent Claim
134 words
An enzyme-linked immunosorbent assay (ELISA) for diagnosing human infection caused by HSV-1 which comprises: (a) immobilizing a target antigen which is a mixture of monoclonal antibody immunoaffinity purifed HSV-1 gC and gD as HSV-1 antigen; (b) absorbing human test serum with HSV-2-infected cell culture extract; (c) reacting said target antigen with the human test serum of step "b" to form a first antigen-antibody complex; (d) reacting said first complex with an enzyme-linked anti-human immunoglobulin to form a second complex; (e) reacting said second complex with a color-producing solution specific for the enzyme in step "c"; (f) repeating steps b-e with a second portion of test serum and extract from non-infected cell culture; and (g) measuring the ratio of absorbance generated by the human serum of step "b" to the human serum of step "f".
Claim 9
Dependent Claim
13 words
The assay of claim 5 wherein said anti-human immuoglobulin is made in goats.
Claim 10
Dependent Claim
129 words
An enzyme-linked immunosorbent assay (ELISA) for diagonosing human infection caused by HSV-2 which comprises: (a) immobilizing a target antigen which is a mixture of monoclonal antibody immunoaffinity purified HSV-2 gD and gF as HSV-2 antigens; (b) absorbing human test serum with HSV-1-infected cells; (c) reacting said target antigen with the human test serum of step "b" to form a first antigen-antibody complex; (d) reacting said first complex with an enzyme-linked anti-human immunoglobulin to form a second complex; (e) reacting said second complex with a color-producing substraste specific for the enzyme in step "c"; (f) repeating steps b-e with a second portion of test serum and uninfected cells; and (g) measuring the ratio of absorbance generated by the human serum of step "b" to the human serum of step "f."
Claim 11
Dependent Claim
129 words
An enzyme-linked immunosorbent assay (ELISA) for diagonosing human infection caused by HSV-2 which comprises: (a) immobilizing a target antigen which is a mixture of monoclonal antibody immunoaffinity purified HSV-2 gD and gF as HSV-2 antigens; (b) absorbing human test serum with HSV-1-infected cells; (c) reacting said target antigen with the human test serum of step "b" to form a first antigen-antibody complex; (d) reacting said first complex with an enzyme-linked anti-human immunoglobulin to form a second complex; (e) reacting said second complex with a color-producing substraste specific for the enzyme in step "c"; (f) repeating steps b-e with a second portion of test serum and uninfected cells; and (g) measuring the ratio of absorbance generated by the human serum of step "b" to the human serum of step "f."
Claim 12
Dependent Claim
13 words
The assay of claim 7 wherein said anti-human immunoglobulin is made in goats.
Claim 13
Dependent Claim
142 words
A test kit for assaying antibodies directed against HSV-1 or HSV-2 in human sera which comprises (a) microtiter plates coated with purified glycoprotein mixtures encoded by HSV-1 or HSV-2 (target antigens) and treated with a protein solution to prevent further absorption of proteins to the plates by means other than antigen-antibody interaction; (b) control positive human serum of known reactivity and antibody titer to HSV-1 or HSV-2 target antigens; (c) control negative human serum lacking antibodies to HSV-1 or HSV-2 target antigens; (d) uninfected Viro cell extracts for absorption of said human serum; (e) HSV-1 infected or HSV-2 infected Viro cell extracts for absorption of said human serum; (f) enzyme-conjugated anti-human immunoglobulin; (g) color-producing substrate solution for use as a substrate for the enzyme of step f; (h) a solution to stop the enzyme reaction; and (i) washing buffers and solvent buffers.
Claim 14
Dependent Claim
142 words
A test kit for assaying antibodies directed against HSV-1 or HSV-2 in human sera which comprises (a) microtiter plates coated with purified glycoprotein mixtures encoded by HSV-1 or HSV-2 (target antigens) and treated with a protein solution to prevent further absorption of proteins to the plates by means other than antigen-antibody interaction; (b) control positive human serum of known reactivity and antibody titer to HSV-1 or HSV-2 target antigens; (c) control negative human serum lacking antibodies to HSV-1 or HSV-2 target antigens; (d) uninfected Viro cell extracts for absorption of said human serum; (e) HSV-1 infected or HSV-2 infected Viro cell extracts for absorption of said human serum; (f) enzyme-conjugated anti-human immunoglobulin; (g) color-producing substrate solution for use as a substrate for the enzyme of step f; (h) a solution to stop the enzyme reaction; and (i) washing buffers and solvent buffers.
References Cited (Backward Citations)
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1991-08-16 | 1992-02-15 | 1992-08-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-08-16 | 1996-02-15 | 1996-08-16 | $3760.00 | Window Closed / Lapsed |
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