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US 5,183,735 AGranted
Method and diagnostic test kit for detection of anti-dsDNA antibodies
AssigneeReaads Medical Products, Inc.
InventorsLuis R. Lopez, Marcia Sterhan
Life of this patent
Estimating the term…
Abstract
A diagnostic element for detecting the presence of anti-dsDNA antibodies in a sample. The diagnostic element includes a support (e.g., a microcell), a coating of mBSA on the support, dsDNA antigens immobilized on the coating, and a blocking layer for blocking immobilization sites other than ones occupied by dsDNA antigens. A testing kit is also provided which includes a diagnostic element, a sample diluent, an antibody solution, and a label identifying solution.
Claims13
A diagnostic element adapted to detect the presence of anti-dsDNA antibodies in a sample specimen upon contact with said sample specimen, comprising:
- a support member, selected from the group consisting of plates, test tubes, and wells;
- a coating on a surface portion of said support member, said first coating consisting essentially of methylated bovine serum albumin as a constituent thereof whereby said methylated bovine serum albumin provides immobilization sites;
- dsDNA antigens specific for said anti-dsDNA antibodies, said dsDNA antigens being immobilized on said coating at occupied ones of said immobilization sites;
- and
- a blocking layer disposed on said coating, said blocking layer consisting essentially of a blocking material operative to block a majority of said immobilization sites other than the occupied ones thereof whereby said blocking layer provides a contact surface available to be contacted by said sample specimen with said contact surface having dsDNA antigens forming binding sites specific for said anti-dsDNA antibodies while being substantially free of ssDNA antigens forming binding sites;
- wherein said blocking layer is operative to stabilize said dsDNA antigens immobilized on the occupied ones of said immobilization sites;
- and wherein said contact surface has a specificity greater than sixty-two percent.
4 more elements
A test kit for detecting the presence of anti-dsDNA specific antibodies in a sample specimen, comprising:
- (a) a diagnostic element including a support member, selected from the group consisting of plates, test tubes and wells having a coating consisting essentially of methylated bovine serum albumin that provides immobilization sites, dsDNA antigens attached at occupied ones of said immobilization sites, said dsDNA antigens specific for said anti-dsDNA antibodies, and a blocking layer consisting essentially of a blocking material operative to block a majority of said immobilization sites other than the occupied ones thereof, said diagnostic element thereby providing a contact surface having dsDNA antigens forming binding sites specific for said anti-dsDNA specific antibodies whereby said anti-dsDNA specific antibodies may be bound to said diagnostic element;
- (b) a sample diluent for use in diluting said sample specimen prior to application thereof to said contact surface;
- (c) a conjugated antibody solution including anti-human IgM and IgG antibodies conjugated to a label and operative to affix to bound ones of said anti-dsDNA specific antibodies upon contact therewith such that the relative amount of anti-dsDNA specific antibodies in said sample specimens can be determined;
- (d) a label identifying solution operative to permit analyzation of attached conjugated antibody solution labels.
1 more element
A method of producing a diagnostic element adapted to detect the presence of anti-dsDNA antibodies in a sample specimen, comprising the steps of:
- providing a support selected from the group consisting of plates, test tubes, and wells;
- coating a surface portion of said support with a first substance consisting essentially of methylated bovine serum albumin to form a layer having immobilization sites;
- contacting said layer with dsDNA antigens specific for anti-dsDNA antibodies whereby at least some of said dsDNA antigens attach to said methylated bovine serum albumin at some of said immobilization sites;
- washing said first layer after attachment of said dsDNA antigens with a second substance including an enzyme capable of digesting ssDNA;
- and
- coating said first layer with a third substance consisting essentially of a blocking material operative to eliminate available ones of said immobilization sites and to stabilize said dsDNA antigens at a contact surface;
- wherein said contact surface has a specificity greater than sixty-two percent.
4 more elements
A method for detecting the presence of anti-dsDNA antibody in a sample specimen comprising the steps of:
- (a) providing a diagnostic element in the form of a support member, selected from the group consisting of plates, test tubes, and wells having a coating comprising consisting essentially of methylated bovine serum albumin that provides immobilization sites, dsDNA antigens attached at occupied ones of said immobilization sites, said dsDNA antigens specific for said anti-dsDNA antibodies, and a blocking layer consisting essentially of a blocking material operative to block a majority of said immobilization sites other than the occupied ones thereof, said diagnostic element thereby providing a contact surface having dsDNA antigens forming binding sites specific for said anti-dsDNA specific antibodies whereby said anti-dsDNA specific antibodies may be bound to said diagnostic element;
- (b) contacting said contact surface with said sample specimen to be tested for the presence therein of said anti-dsDNA antibody;
- (c) within 15 minutes after step (b), contacting said contact surface with a labeled antibody whereby at least some of said labeled antibody binds to said anti-dsDNA antibody;
- (d) rinsing the contact surface after contact thereof with said labeled antibody;
- and
- (e) measuring the amount of said bound label within 105 minutes after contact of said contact surface with said sample specimen whereby an amount of anti-dsDNA antibody may be determined.
3 more elements
Dependencies are inferred from the claim text.