Patent US5030561A successfully refreshed from Google Patents.
US5030561A
Expired / Lapsed
US
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Chlamydia assay using amidine modified supports or particles
Assignee: Becton Dickinson and Co
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Filing: 1988-12-27
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Granted: 1991-07-09
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Expires: 2008-12-27
Abstract
Abstract A method for assaying of Chlamydia includes adhering Chlamydia antigen to amidine modified latex particles, binding of adhered antigen to an anti-Chlamydia antibody conjugated to an enzyme, separating the particles from the liquid phase of the assay and detecting bound enzyme by color development when the separated particles are contacted with a substrate for the enzyme. The invention includes a kit of materials for preforming an assay in accordance with the method of the invention.
Inventors
- 👤 James P. Mapes
- 👤 Catherine S. Donahue
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 21:41
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; G01N33/56911; G01N33/56927; G01N33/543; G01N33/54313; Y; Y10; Y10S; Y10S436/00; Y10S436/823
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
122 words
A method for determining Chlamydia antigen comprising: a) contacting particles of an amidine modified latex with a liquid suspected of containing Chlamydia antigen whereby said antigen adheres to said particles; b) contacting said particles having antigen adhered thereto with an anti-Chlamydia antibody conjugated to an enzyme whereby said antigen on said particles binds to said antibody to give a bound fraction containing said enzyme on said particles; c) separating said particles on said bound fraction from said liquid; d) contacting the enzyme on said separated particles with a substrate reactive with said enzyme whereby said enzyme converts said substrate to a product; and e) detecting the presence of Chlamydia antigen in said sample by the appearance of color associated with said product.
Claim 2
Independent Claim
122 words
A method for determining Chlamydia antigen comprising: a) contacting particles of an amidine modified latex with a liquid suspected of containing Chlamydia antigen whereby said antigen adheres to said particles; b) contacting said particles having antigen adhered thereto with an anti-Chlamydia antibody conjugated to an enzyme whereby said antigen on said particles binds to said antibody to give a bound fraction containing said enzyme on said particles; c) separating said particles on said bound fraction from said liquid; d) contacting the enzyme on said separated particles with a substrate reactive with said enzyme whereby said enzyme converts said substrate to a product; and e) detecting the presence of Chlamydia antigen in said sample by the appearance of color associated with said product.
Claim 3
Dependent Claim
37 words
The method of claim 1 further comprising determining the concentration of said Chlamydia antigen in said liquid by comparing the magnitude of said color with the magnitude of color established for a known quantity of the antigen.
Claim 4
Dependent Claim
21 words
The method of claim 1 wherein said latex is selected from the group consisting of polystyrene, styrene butadiene copolymer and polyacrylate.
Claim 5
Dependent Claim
20 words
The method of claim 1 wherein said enzyme is selected from the group consisting of a peroxidase and a hydrolase.
Claim 6
Dependent Claim
20 words
The method of claim 4 wherein said hydrolase is selected from the group consisting of alkaline phosphatase, galactosidase and carboxyesterase.
Claim 7
Dependent Claim
12 words
The method of claim 1 wherein said antibody is a monoclonal antibody.
Claim 8
Dependent Claim
12 words
The method of claim 1 wherein said separating is performed by centrifugation.
Claim 9
Dependent Claim
15 words
The method of claim 1 wherein said separating is performed by filtration through a membrane.
Claim 10
Dependent Claim
14 words
The method of claim 8 wherein said membrane is pretreated with an inert protein.
Claim 11
Dependent Claim
19 words
The method of claim 1 wherein the enzyme on the separated particles is additionally contacted with a tetrazolium salt.
Claim 12
Dependent Claim
11 words
The method of claim 10 wherein said salt is nitroblue tetrazolium.
Claim 13
Independent Claim
71 words
A method for determining Chlamydia antigen comprising: a) adhering Chlamydia antigen in a liquid to an amidine derivatized solid support; b) contacting antigen adhered to said support with an antibody having a label conjugated thereto, said antigen binding to said antibody to give a bound fraction containing said label on said support; c) separating said support from said liquid; and d) detecting said antigen by a signal associated with said label.
Claim 14
Independent Claim
71 words
A method for determining Chlamydia antigen comprising: a) adhering Chlamydia antigen in a liquid to an amidine derivatized solid support; b) contacting antigen adhered to said support with an antibody having a label conjugated thereto, said antigen binding to said antibody to give a bound fraction containing said label on said support; c) separating said support from said liquid; and d) detecting said antigen by a signal associated with said label.
Claim 15
Dependent Claim
24 words
The method of claim 12 wherein said label is selected from the group consisting of a radioactive atom, a fluorescent dye and an enzyme.
Claim 16
Dependent Claim
25 words
The method of claim 13 wherein said label is a radioactive atom and said detecting is performed by measuring radioactive counts from said bound fraction.
Claim 17
Dependent Claim
32 words
The method of claim 13 wherein said label is a fluorescent dye, and said detecting is performed by subjecting said bound fraction to excitation light and measuring fluorescence from said bound fraction.
Claim 18
Independent Claim
128 words
A method for determining Chlamydia organisms in a liquid comprising: a) subjecting a liquid suspected of containing Chlamydia organisms to a reagent for exposing Chlamydia antigen; b) contacting said liquid with a latex containing amidine modified particles whereby said antigen adheres to said particles; c) contacting said particles having adhered antigen with a monoclonal antibody conjugated to alkaline phosphatase whereby said antigen adhered to said particles binds to said antibody to give a bound fraction containing alkaline phosphatase on said particles; d) filtering said latex through a membrane having a pore size sufficient to retain said particles; e) adding a substrate reactive with said alkaline phosphatase to said membrane; and f) detecting the presence of Chlamydia organisms in said liquid by the appearance of color on said membrane.
Claim 19
Independent Claim
128 words
A method for determining Chlamydia organisms in a liquid comprising: a) subjecting a liquid suspected of containing Chlamydia organisms to a reagent for exposing Chlamydia antigen; b) contacting said liquid with a latex containing amidine modified particles whereby said antigen adheres to said particles; c) contacting said particles having adhered antigen with a monoclonal antibody conjugated to alkaline phosphatase whereby said antigen adhered to said particles binds to said antibody to give a bound fraction containing alkaline phosphatase on said particles; d) filtering said latex through a membrane having a pore size sufficient to retain said particles; e) adding a substrate reactive with said alkaline phosphatase to said membrane; and f) detecting the presence of Chlamydia organisms in said liquid by the appearance of color on said membrane.
Claim 20
Dependent Claim
11 words
The method of claim 16 wherein said reagent contains a detergent.
Claim 21
Dependent Claim
39 words
A kit of materials for performing an assay for Chlamydia comprising a latex of amidine modified polystyrene particles, an anti Chlamydia antibody conjugated to an enzyme, a substrate reactive with said enzyme, and a membrane for filtering said particles.
Claim 22
Dependent Claim
39 words
A kit of materials for performing an assay for Chlamydia comprising a latex of amidine modified polystyrene particles, an anti Chlamydia antibody conjugated to an enzyme, a substrate reactive with said enzyme, and a membrane for filtering said particles.
Claim 23
Dependent Claim
13 words
The kit of claim 18 further comprising a reagent for altering Chlamydia organisms.
Claim 24
Dependent Claim
18 words
The kit of claim 18 further comprising one or more solutions containing a predetermined quantity of Chlamydia antigen.
Claim 25
Dependent Claim
19 words
The kit of claim 20 further comprising a solution or utensil useful in performing the assay of the invention.
Claim 26
Dependent Claim
27 words
The kit of claim 18 further comprising a housing for assembly of the components of said kit, said housing containing an absorbent material positioned under said membrane.
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) | 1994-07-08 | 1995-01-07 | 1995-07-09 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-07-08 | 1999-01-07 | 1999-07-09 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-07-08 | 2003-01-07 | 2003-07-09 | $7700.00 | Window Closed / Lapsed |