EP0174106A2
Expired / Lapsed
EP
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Detection of cell membrane protein
Assignee: Becton Dickinson and Co
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Filing: 1985-08-08
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Granted: —
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Expires: 2005-08-08
Abstract
Abstract The invention comprises a method for detecting cell proteins of microorganisms, such as the principal outer membrane protein of Chlamydia trachomatis having a mean molecular weight of 39.5 daltons. The method includes the steps of adding a buffer salt solution to a specimen suspected of containing bacterial antigens, raising the pH of the buffered solution so produced, incubating the solution, adding a neutralizing buffer to the solution to lower the pH, and assaying the sample by conventional immunoassay techniques. Optionally the sample solution is heated prior to incubation and then cooled afterwards before adding the neutralizing buffer.
Inventors
- 👤 Larry Mosier
- 👤 John Petersen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 07:55
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; G01N33/56911; G01N33/56927
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
110 words
1 A method for detecting cell membrane proteins of microorganisms, the method comprising: taking a test sample; forming a sample solution by mixing the sample with a first buffer solution having a pH of from 6 to 8: adjusting the sample solution pH to a value of from 8 to 12.5 using a solution of base; incubating the sample for a period of from 5 minutes to 30 minutes; adding a neutralizing second buffer having a pH of from 1 to 7 to bring the pH of the sample solution to a final value of from 7 to 8; and assaying the sample solution to detect the presence of antigens.
Claim 2
Dependent Claim
110 words
1 A method for detecting cell membrane proteins of microorganisms, the method comprising: taking a test sample; forming a sample solution by mixing the sample with a first buffer solution having a pH of from 6 to 8: adjusting the sample solution pH to a value of from 8 to 12.5 using a solution of base; incubating the sample for a period of from 5 minutes to 30 minutes; adding a neutralizing second buffer having a pH of from 1 to 7 to bring the pH of the sample solution to a final value of from 7 to 8; and assaying the sample solution to detect the presence of antigens.
Claim 3
Dependent Claim
18 words
A method according to claim 1 wherein the first buffer is a sugar-containing buffer solution, especially sucrose phosphate.
Claim 4
Dependent Claim
20 words
A method according to claim 1 or 2 wherein the base comprises sodium hydroxide, potassium hydroxide, trisodium phosphate, or tri(hydroxymethyl)aminomethane.
Claim 5
Dependent Claim
52 words
A method according to claim 1. 2 or 3 wherein the incubating step includes heating the sample solution to a temperature of from 20°C to 105°C after adjusting the sample pH, incubating the sample at that temperature, and cooling the sample solution after incubation to from 0°C to 40°C, preferably about 25°C.
Claim 6
Dependent Claim
20 words
A method according to claim 4 wherein the sample solution is heated to a temperature of from 90°C to 100°C
Claim 7
Independent Claim
25 words
A method according to any preceding claim wherein the neutralizing second buffer comprises a phosphate, citric acid, hydrochloric acid or tri(hydroxymethyl)aminomethane.HCl solution, especially sucrose phospate.
Claim 8
Independent Claim
20 words
A method according to any preceding claim, wherein the final pH of the sample solution is from 7.2 to 7.8.
Claim 9
Independent Claim
29 words
A method according to any preceding claim, wherein after mixing the sample with a first buffer solution, the pH of the sample is adjusted to between 11.0 and 11.8.
Claim 10
Dependent Claim
14 words
A method according to claim 8, wherein the pH is adjusted with sodium hydroxide.
Claim 11
Independent Claim
13 words
A method according to any preceding claim, when used to detect Chlamydia trachomatis.
Claim 12
Independent Claim
13 words
A method according to any preceding claim, wherein the assaying is by radioimmunoassay.
Claim 13
Independent Claim
19 words
A method according to any of claims 1 to 10, wherein the assaying is by enzyme linked immunoabsorbent assay.
Claim 14
Independent Claim
158 words
A method for detecting the principal outer membrane protein of Chlam y dia trachomatis, the method comprising: taking a sample such as a cervical or urethral test swab; placing the sample in a buffer solution comprising sucrose phosphate having a pH of about 7.0: mixing the sample with the buffer solution to form a sample solution: adding to the sample solution a quantity of sodium hydroxide solution having a molarity of about 0.4 thereby to raise the pH of the sample solution to from 11.0 to 11.8; raising the temperature of the sample solution to about 100°C; incubating the sample solution at about 100°C for a period of about 15 minutes: cooling the sample solution to a temperature of from 20°C to 30°C; reducing the pH of the cooled sample solution to between 7.2 and 7.8, using a neutralizing buffer of phosphate having a pH of about 6.1; and assaying the sample to detect the presence of antigen.
Claim 15
Independent Claim
158 words
A method for detecting the principal outer membrane protein of Chlam y dia trachomatis, the method comprising: taking a sample such as a cervical or urethral test swab; placing the sample in a buffer solution comprising sucrose phosphate having a pH of about 7.0: mixing the sample with the buffer solution to form a sample solution: adding to the sample solution a quantity of sodium hydroxide solution having a molarity of about 0.4 thereby to raise the pH of the sample solution to from 11.0 to 11.8; raising the temperature of the sample solution to about 100°C; incubating the sample solution at about 100°C for a period of about 15 minutes: cooling the sample solution to a temperature of from 20°C to 30°C; reducing the pH of the cooled sample solution to between 7.2 and 7.8, using a neutralizing buffer of phosphate having a pH of about 6.1; and assaying the sample to detect the presence of antigen.
References Cited (Backward Citations)
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