Method for screening for cardiomyopathy

• US 5,418,139 A
• Filed: 02/10/1993
• Issued: 05/23/1995
• Est. Priority Date: 02/10/1993
• Status: Expired due to Term

• Current Assignee: University of Iowa Research Foundation
• Original Assignee: University of Iowa Research Foundation
• Inventors: Campbell, Kevin P.
• Primary Examiner(s): Scheiner, Toni R.
• Assistant Examiner(s): Parsons, Nancy J.
• Application Number:
  US08/016,126
• Time in Patent Office: 832 Days
• Field of Search: 435/7.21, 435/960, 436/518, 436/536, 436/172, 436/811
• US Class Current: 435/7.21
• CPC Class Codes: G01N 2474/20 Immunohistochemistry assay
• CPC Class Codes: G01N 2800/325 Heart failure or cardiac arrest, e.g. cardiomyopathy, congestive heart failure
• CPC Class Codes: G01N 33/6887 from muscle, cartilage or connective tissue
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function

Abstract

Disclosed herein are methods for screening for primary cardiomyopathy. The methods are preferably immunological methods in which the level of binding of a monoclonal or polyclonal antibody to a 50 kD glycoprotein component of a mammalian muscle tissue is determined. This level of binding is compared to the level of binding observed when non-dystrophic tissue is treated in an otherwise identical manner. A substantial reduction in the level of binding to the 50 kD glycoprotein in the experimental mammalian muscle tissue has been determined to be a screen for primary cardiomyopathy.

Claims

1. A method for screening for primary cardiomyopathy in a mammal comprising the steps of:

a) providing a muscle tissue sample from the mammal;

b) contacting the muscle tissue sample, or a component thereof, with an antibody which binds to a dystrophin-associated glycoprotein having a molecular weight of about 50 kD, under conditions appropriate for binding;

c) detecting the level of binding of the antibody to the 50 kD dystrophin-associated glycoprotein from the muscle tissue;

d) comparing the level of binding of the antibody to the 50 kD dystrophin-associated glycoprotein from the muscle tissue, to the level of binding of the antibody to the 50 kD dystrophin-associated glycoprotein in a normal control muscle tissue, a substantial reduction in the level of binding in the tissue, as compared to control tissue, being indicative of primary cardiomyopathy.

2. A method of claim 1 wherein the experimental muscle tissue sample is a skeletal muscle tissue sample or a cardiac muscle tissue sample.

3. A method of claim 2 wherein the mammal is a human.

4. A method for screening for primary congestive cardiomyopathy in a mammal comprising the steps of:

a) providing a cardiac muscle tissue biopsy sample suitable for histochemical analysis;

b) contacting the cardiac muscle tissue biopsy sample with an antibody which binds to a dystrophin-associated glycoprotein having a molecular weight of about 50 kD;

c) washing the cardiac muscle tissue biopsy sample to remove non-specifically bound antibody;

d) detecting the level of binding of the antibody specific for the 50 kD dystrophin-associated glycoprotein from the cardiac muscle tissue biopsy sample; and

e) comparing the level of binding of the antibody to the 50 kD dystrophin-associated glycoprotein from the cardiac muscle tissue biopsy sample, to the level of binding of the antibody specific for the 50 kD dystrophin-associated glycoprotein in a normal control cardiac tissue biopsy sample, a substantial reduction in the level of binding in the tissue, as compared to control tissue, being indicative of primary congestive cardiomyopathy.

5. A method of claim 4 wherein the mammal is a human.

6. A method of claim 5 wherein the antibody is detectable by immunofluorescence microscopy.

7. A method for screening for primary congestive cardiomyopathy in a mammal comprising the steps of:

a) providing solubilized muscle cell extracts or membranes from the mammal;

b) separating the components of the solubilized muscle cell membranes by gel electrophoresis;

c) transferring the separated components from step b) to a solid support;

d) contacting the components of the solubilized muscle cell membranes from step c) with an antibody which binds to a dystrophin-associated glycoprotein having a molecular weight of about 50 kD, under conditions appropriate for binding of the antibody to the dystrophin-associated protein;

e) washing the components of the solubilized muscle cell membranes from step d) to remove non-specifically bound antibody;

f) detecting the level of binding of the antibody to the 50 kD dystrophin-associated glycoprotein from the muscle cell membranes; and

g) comparing the level of binding of the antibody to the 50 kD dystrophin-associated glycoprotein from the muscle cell to the level of binding of the antibody to the 50 kD dystrophin-associated glycoprotein from a normal control muscle cell membrane treated as in steps a) through f), a substantial reduction in the level of binding in the muscle cell membranes, as compared to control muscle cell membranes, being indicative of primary congestive cardiomyopathy.

8. A method of claim 7 wherein the muscle cell extracts or membranes are from skeletal muscle tissue.

9. A method of claim 8 wherein the mammal is a human.

10. A method of claim 7 wherein the muscle cell extracts or membranes are from cardiac muscle tissue.