US4248964A
Expired / Lapsed
US
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Detection and quantitation of Neisseria via radioimmunoassay of an enzyme present in Neisseria bacteria
Assignee: Bayer Corp
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Filing: 1977-09-28
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Granted: 1981-02-03
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Expires: 1997-09-28
Abstract
Abstract This invention relates to a method for detecting Neisseria bacteria by means of a radioimmunoassay for a bacterial protein. More specifically, this invention employs the method of radioimmunoassay in which a purified enzyme or enzyme related protein from Neisseria cells is labelled and is mixed with the unknown sample which represents a bacterial lysate. A known amount of antibody specific for the enzyme is added to the mixture and forms complexes with the labelled and unlabelled enzyme present. After a suitable reaction period, the labelled enzyme-antibody complex is separated from the mixture, and its radioactivity is determined. The presence and amount of enzyme in the unknown sample is calculated by relating the radioactivity count to a standard curve prepared with known quantities of unlabelled enzyme.
Inventors
- 👤 Hugh C. McDonald
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 19:43
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; C12Q1/12; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; G01N33/571; Y; Y10; Y10S; Y10S436/00; Y10S436/804
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
91 words
A method for detecting Neisseria bacteria in a fluid sample via a radioimmunoassay technique consisting of the steps: (1) bringing radiolabelled 1,2-propanediol dehydrogenase or its subunit into contact with a lysate of the sample to be tested; (2) bringing an antiserum specific to 1,2-propanediol dehydrogenase into contact with the above and incubating the mixture; (3) separating the unlabelled and radiolabelled 1,2-propanediol dehydrogenase-antibody complexes from the uncomplexed reactants in said mixture; and then (4) monitoring either or both portions of the mixture for the presence of radiolabelled 1,2-propanediol dehydrogenase or its subunit.
Claim 2
Independent Claim
91 words
A method for detecting Neisseria bacteria in a fluid sample via a radioimmunoassay technique consisting of the steps: (1) bringing radiolabelled 1,2-propanediol dehydrogenase or its subunit into contact with a lysate of the sample to be tested; (2) bringing an antiserum specific to 1,2-propanediol dehydrogenase into contact with the above and incubating the mixture; (3) separating the unlabelled and radiolabelled 1,2-propanediol dehydrogenase-antibody complexes from the uncomplexed reactants in said mixture; and then (4) monitoring either or both portions of the mixture for the presence of radiolabelled 1,2-propanediol dehydrogenase or its subunit.
Claim 3
Dependent Claim
17 words
A method according to claim 1 wherein said fluid sample is a human body fluid or exudate.
Claim 4
Dependent Claim
19 words
A method according to claim 1 wherein lysis and incubation are carried out at a pH between about 7-10.
Claim 5
Dependent Claim
19 words
A method according to claim 1 wherein lysis and incubation are carried out at temperatures between about 0°-60° C.
Claim 6
Dependent Claim
20 words
A method according to claim 1 wherein said antiserum is immobilized on a substrate which is inert toward the reactants.
Claim 7
Dependent Claim
20 words
A method according to claim 5 wherein said antiserum is immobilized on said substrate via physical adsorption or covalent bonding.
Claim 8
Dependent Claim
16 words
A method according to claim 1 wherein said Neisseria bacteria are of the species Neisseria gonorrhoeae.
Claim 9
Independent Claim
114 words
A method for detecting Neisseria bacteria in a fluid sample via a radioimmunoassay technique consisting of the steps: (1) preparing an antiserum specific to 1,2-propanediol dehydrogenase; (2) purifying 1,2-propanediol dehydrogenase or its subunit; (3) preparing radiolabelled 1,2-propanediol dehydrogenase or its subunit; (4) preparing a lysate of said fluid sample; (5) mixing and incubating said lysate with said antiserum and said radiolabelled and unlabelled 1,2-propanediol dehydrogenase or its subunit; (6) admixing an antiglobulin reagent which causes a precipitation of the 1,2-propanediol dehydrogenase-antibody complex or 1,2-propanediol dehydrogenase subunit-antibody complex, thereby separating them from the uncomplexed reagents; and then (7) monitoring the precipitate and/or the supernatant liquid for the presence of radiolabelled 1,2-propanediol dehydrogenase or its subunit.
Claim 10
Independent Claim
114 words
A method for detecting Neisseria bacteria in a fluid sample via a radioimmunoassay technique consisting of the steps: (1) preparing an antiserum specific to 1,2-propanediol dehydrogenase; (2) purifying 1,2-propanediol dehydrogenase or its subunit; (3) preparing radiolabelled 1,2-propanediol dehydrogenase or its subunit; (4) preparing a lysate of said fluid sample; (5) mixing and incubating said lysate with said antiserum and said radiolabelled and unlabelled 1,2-propanediol dehydrogenase or its subunit; (6) admixing an antiglobulin reagent which causes a precipitation of the 1,2-propanediol dehydrogenase-antibody complex or 1,2-propanediol dehydrogenase subunit-antibody complex, thereby separating them from the uncomplexed reagents; and then (7) monitoring the precipitate and/or the supernatant liquid for the presence of radiolabelled 1,2-propanediol dehydrogenase or its subunit.
Claim 11
Dependent Claim
17 words
A method according to claim 8 wherein said fluid sample is a human body fluid or exudate.
Claim 12
Dependent Claim
19 words
A method according to claim 8 wherein lysis and incubation are carried out at a pH between about 7-10.
Claim 13
Dependent Claim
19 words
A method according to claim 8 wherein lysis and incubation are carried out at temperatures between about 0°-60° C.
Claim 14
Dependent Claim
20 words
A method according to claim 8 wherein said antiserum is immobilized on a substrate which is inert toward the reactants.
Claim 15
Dependent Claim
20 words
A method according to claim 12 wherein said antiserum is immobilized on said substrate via physical adsorption or covalent bonding.
Claim 16
Dependent Claim
16 words
A method according to claim 8 wherein said Neisseria bacteria are of the species Neisseria gonorrhoeae.
References Cited (Backward Citations)
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