METHOD AND KIT FOR THE DETECTION OF HEPATITIS-SPECIFIC ANTIBODIES
First Claim
1. A method for the detection of hepatitis virus-specific antibodies in a sample from a subject comprising the steps of:
- a) incubating a sample from said subject in a culture in the presence of a medium comprising a) one or more activators of (i) hepatitis virus-primed lymphocytes, (ii) memory cells specific for said hepatitis virus, (iii) hepatitis virus-specific antibody production, or (iv) a combination thereof, and b) an antibody against a B-lymphocyte membrane domain;
b) exposing the resultant culture of step a) to a hepatitis virus antigen, thereby allowing an antigen-antibody immune complex to form; and
c) detecting the antigen-antibody immune complex of step b);
thereby detecting the presence of hepatitis virus-specific antibodies.
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Accused Products
Abstract
The present invention relates to an improved method for detecting antibodies to a hepatitis virus in a tissue sample from individuals, which can reliably detect antibodies in recently infected individuals and which provides much lower false positive results in individuals that have cleared their hepatitis infections. More particularly, the present invention relates to an improved method and kit which utilizes an activator of (i) hepatitis virus-primed lymphocytes, (ii) memory cells specific for said hepatitis virus, (iii) hepatitis virus-specific antibody production, or (iv) a combination thereof in a tissue sample to stimulate the production of antibodies from newly primed B cells, if present.
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Citations
141 Claims
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1. A method for the detection of hepatitis virus-specific antibodies in a sample from a subject comprising the steps of:
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a) incubating a sample from said subject in a culture in the presence of a medium comprising a) one or more activators of (i) hepatitis virus-primed lymphocytes, (ii) memory cells specific for said hepatitis virus, (iii) hepatitis virus-specific antibody production, or (iv) a combination thereof, and b) an antibody against a B-lymphocyte membrane domain; b) exposing the resultant culture of step a) to a hepatitis virus antigen, thereby allowing an antigen-antibody immune complex to form; and c) detecting the antigen-antibody immune complex of step b); thereby detecting the presence of hepatitis virus-specific antibodies. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 32, 33, 34, 35, 36, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 53, 54, 55, 56, 57, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 77, 80, 81, 83, 84, 85, 86, 87, 88, 89, 90, 91, 97, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 117, 118, 119, 120, 121, 122, 123, 124, 127, 128, 129, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141)
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26. A method for the detection of a hepatitis infection in a subject comprising the steps of:
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a. incubating a sample from said subject in a culture in the presence of a medium comprising a) one or more activators of (i) hepatitis virus-primed lymphocytes, (ii) memory cells specific for said hepatitis virus, (iii) hepatitis virus-specific antibody production, or (iv) a combination thereof, and b) an antibody against a B-lymphocyte membrane domain; b. exposing the resultant culture of step a) to a hepatitis virus antigen, thereby allowing an antigen-antibody immune complex to form; and c. detecting the antigen-antibody immune complex of step b); thereby detecting a hepatitis infection in said subject. - View Dependent Claims (27, 28, 29, 30, 31, 37, 38)
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51. A method for the detection of a very early hepatitis infection in a subject comprising the steps of:
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a. incubating a sample from said subject in a culture in the presence of a medium comprising a) one or more activators of (i) hepatitis virus-primed lymphocytes, (ii) memory cells specific for said hepatitis virus, (iii) hepatitis virus-specific antibody production, or (iv) a combination thereof, and b) an antibody against a B-lymphocyte membrane domain; b. exposing the resultant culture of step a) to a hepatitis virus antigen, thereby allowing an antigen-antibody immune complex to form; and c. detecting the antigen-antibody immune complex of step b); thereby detecting a very early hepatitis infection in said subject. - View Dependent Claims (52, 58, 59)
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72. A method for detecting a hepatitis infection in a seronegative subject comprising the steps of:
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a. incubating a sample from said subject in a culture in the presence of a medium comprising a) one or more activators of (i) hepatitis virus-primed lymphocytes, (ii) memory cells specific for said hepatitis virus, (iii) hepatitis virus-specific antibody production, or (iv) a combination thereof, and b) an antibody against a B-lymphocyte membrane domain; b. exposing the resultant culture of step a) to a hepatitis virus antigen, thereby allowing an antigen-antibody immune complex to form; and c. detecting the antigen-antibody immune complex of step b); wherein if the antigen-antibody immune complex is detected, said seronegative subject has a hepatitis infection, thereby detecting a hepatitis infection in said seronegative subject. - View Dependent Claims (73, 74, 75, 76, 78, 79, 82)
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92. A method of differentiating between a cleared and chronic hepatitis infection in a subject comprising the steps of:
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a. obtaining a tissue sample from said subject; b. determining the anti-hepatitis virus antibody level in a first aliquot of said sample, wherein a detectable anti-hepatitis virus antibody level indicates that a subject is seropositive; c. stimulating a second aliquot of said sample to produce anti-hepatitis virus antibodies in vitro by incubating said second aliquot in a culture in the presence of a medium comprising a) one or more activators of (i) hepatitis virus-primed lymphocytes, (ii) memory cells specific for said hepatitis virus, (iii) hepatitis virus-specific antibody production, or (iv) a combination thereof, and b) an antibody against a B-lymphocyte membrane domain; d. determining the anti-hepatitis virus antibody level in said second aliquot of said sample; e. dividing a value representing the stimulated anti-hepatitis virus antibody level obtained in step (d) by a value representing the anti-hepatitis virus antibody level obtained in step (b) for each subject; f. determining if the quotient obtained in step (d) is above a pre-determined threshold value for each subject, wherein a value below said threshold value indicates that the subject has a cleared hepatitis virus infection and a value close to said threshold value indicates that the subject has a chronic hepatitis infection, thereby differentiating between a cleared and chronic hepatitis infection in said subject. - View Dependent Claims (93, 94, 95, 96, 98, 99)
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115. A kit for the detection of hepatitis virus-specific antibodies in a subject comprising:
- a container for retaining a tissue sample, wherein said container comprises a medium, said medium comprising a) one or more activators of (i) lymphocytes activated by said hepatitis virus, (ii) memory cells specific for said hepatitis virus, (iii) antibodies against said hepatitis virus, or (iv) a combination thereof, and b) an antibody against a B-lymphocyte membrane domain.
- View Dependent Claims (116, 125, 126, 130)
Specification