Diagnostic test kit and method for determination of chlamydial or gonococcal antigens using a microporous membrane

Claim 1

A method for the determination of a chlamydial or gonococcal antigen comprising: A. contacting chlamydial or gonococcal antigen extracted from a specimen suspected of containing chlamydial or gonococcal organisms, respectively, said specimen further containing whole blood, mucus or components thereof, with a surfactant-coated polyamide microporous membrane which is substantially free of any particulate matter, and which has an average pore size of from about 1 to about 10 μmeter, said surfactant coating being present in an amount of at least about 20 mg/m 2 , said antigen being immobilized on said coated membrane, B. within about 10 minutes of said contacting step A, contacting chlamydial or gonococcal antigen immobilized on said coated membrane with chlamydial or gonococcal antibody, respectively, so as to form an immunological complex on said membrane, and C. determining the presence of said complex on said membrane as a measure of the presence of chlamydial or gonococcal antigen, respectively, in said specimen.

Claim 2

A method for the determination of a chlamydial or gonococcal antigen comprising: A. contacting chlamydial or gonococcal antigen extracted from a specimen suspected of containing chlamydial or gonococcal organisms, respectively, said specimen further containing whole blood, mucus or components thereof, with a surfactant-coated polyamide microporous membrane which is substantially free of any particulate matter, and which has an average pore size of from about 1 to about 10 μmeter, said surfactant coating being present in an amount of at least about 20 mg/m 2 , said antigen being immobilized on said coated membrane, B. within about 10 minutes of said contacting step A, contacting chlamydial or gonococcal antigen immobilized on said coated membrane with chlamydial or gonococcal antibody, respectively, so as to form an immunological complex on said membrane, and C. determining the presence of said complex on said membrane as a measure of the presence of chlamydial or gonococcal antigen, respectively, in said specimen.

Claim 14

A method for the determination of chlamydial or gonococcal organisms comprising: A. extracting chlamydial or gonococcal antigen from chlamydial or gonococcal organisms, respectively, in a biological specimen containing whole blood, mucus or components thereof, B. contacting said extracted chlamydial or gonococcal antigen with a surfactant-coated polyamide microporous membrane which is substantially free of any particulate matter, and which has an average pore size of from about 1 to about 10 μmeter, said surfactant being present in an amount of at least about 20 mg/m 2 , C. within about 5 minutes of said contacting step B, contacting chlamydial or gonococcal antigen bound to said coated membrane with a reagent composition comprising an unlabeled chlamydial or gonococcal antibody, respectively, so as to form an unlabeled immunological complex on said membrane, D. separating unbound materials in said specimen from said bound complex, E. contacting said bound complex with a labeled antibody to said chlamydial or gonococcal antibody so as to form a labeled immunological complex, F. within about 5 minutes of said contacting step E, separating unbound materials from said labeled complex, and G. determining the presence of said labeled complex on said membrane as a measure of the presence of chlamydial or gonococcal organisms, respectively, in said specimen.

Claim 15

A method for the determination of chlamydial or gonococcal organisms comprising: A. extracting chlamydial or gonococcal antigen from chlamydial or gonococcal organisms, respectively, in a biological specimen containing whole blood, mucus or components thereof, B. contacting said extracted chlamydial or gonococcal antigen with a surfactant-coated polyamide microporous membrane which is substantially free of any particulate matter, and which has an average pore size of from about 1 to about 10 μmeter, said surfactant being present in an amount of at least about 20 mg/m 2 , C. within about 5 minutes of said contacting step B, contacting chlamydial or gonococcal antigen bound to said coated membrane with a reagent composition comprising an unlabeled chlamydial or gonococcal antibody, respectively, so as to form an unlabeled immunological complex on said membrane, D. separating unbound materials in said specimen from said bound complex, E. contacting said bound complex with a labeled antibody to said chlamydial or gonococcal antibody so as to form a labeled immunological complex, F. within about 5 minutes of said contacting step E, separating unbound materials from said labeled complex, and G. determining the presence of said labeled complex on said membrane as a measure of the presence of chlamydial or gonococcal organisms, respectively, in said specimen.

Claim 17

A method for the determination of the lippolysaccharide antigen of C. trachomatis comprising: A. extracting lipopolysaccharide antigen from C. trachomatis organisms in a biological specimen containing whole blood, mucus or components thereof, B. contacting said extracted antigen with a surfactant-coated nylon microporous membrane which is substantially free of any particulate matter, and which has an average pore size of about 5 μmeter, said surfactant being present in an amount of from about 50 to about 150 mg/m 2 , C. separating unbound materials from antigen bound to said coated membrane, and contacting said bound antigen with an unlabeled monoclonal antibody directed to said antigen so as to form an unlabeled immunological complex on said membrane, D. incubating for from about 1 to about 5 minutes at room temperature, E. separating uncomplexed materials from said bound complex by washing, F. contacting said bound complex with a peroxidase-labeled antibody to said chlamydial antibody so as to form a labeled complex bound to said membrane, and incubating for up to about 5 minutes at room temperature, G. separating uncomplexed materials from said bound labeled complex by washing, H. adding a dye-providing composition comprising hydrogen peroxide and a leuco dye which is capable of providing a dye in the presence of peroxidase and hydrogen peroxide, and I. determining the amount of said dye on said membrane as a measure of the amount of C. trachomatis in said specimen.

Claim 18

A method for the determination of the lippolysaccharide antigen of C. trachomatis comprising: A. extracting lipopolysaccharide antigen from C. trachomatis organisms in a biological specimen containing whole blood, mucus or components thereof, B. contacting said extracted antigen with a surfactant-coated nylon microporous membrane which is substantially free of any particulate matter, and which has an average pore size of about 5 μmeter, said surfactant being present in an amount of from about 50 to about 150 mg/m 2 , C. separating unbound materials from antigen bound to said coated membrane, and contacting said bound antigen with an unlabeled monoclonal antibody directed to said antigen so as to form an unlabeled immunological complex on said membrane, D. incubating for from about 1 to about 5 minutes at room temperature, E. separating uncomplexed materials from said bound complex by washing, F. contacting said bound complex with a peroxidase-labeled antibody to said chlamydial antibody so as to form a labeled complex bound to said membrane, and incubating for up to about 5 minutes at room temperature, G. separating uncomplexed materials from said bound labeled complex by washing, H. adding a dye-providing composition comprising hydrogen peroxide and a leuco dye which is capable of providing a dye in the presence of peroxidase and hydrogen peroxide, and I. determining the amount of said dye on said membrane as a measure of the amount of C. trachomatis in said specimen.