Method for the very rapid detection of polyamines

• US 5,330,898 A
• Filed: 10/03/1991
• Issued: 07/19/1994
• Est. Priority Date: 02/20/1991
• Status: Expired due to Term

• Current Assignee: Ampio Pharmaceuticals, Inc.
• Original Assignee: Diagnostic Markers, Inc.
• Inventors: Bar-Or, David, Solomons, Clive
• Primary Examiner(s): Wityshyn, Michael G.
• Assistant Examiner(s): Williams, Jane
• Application Number:
  US07/770,519
• Time in Patent Office: 1,020 Days
• Field of Search: 439/32, 439/29, 439/4, 436/106, 436/111, 436/164, 436/63, 436/811, 436/171
• US Class Current: 435/29
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10T 436/17 Nitrogen containing
• CPC Class Codes: Y10T 436/173845 Amine and quaternary ammonium

Abstract

A method for the detection of at least one polyamine associated with bacterial infection, which comprises contacting a sample suspected of containing polyamine-associated bacteria in aqueous medium at alkaline pH with indigo carmine and dimethyl sulfoxia and observing whether a blue color develops in five minutes indicating the presence of polyamine.

Claims

1. A method for detecting the presence of a polyamine in a sample in which a blue color forms when the polyamine is present and a green color forms when the polyamine is absent, comprising the step of:

contacting an aqueous sample containing a polyamine with a mixture of dimethyl sulfoxide and indigo carmine at an alkaline pH, wherein said blue color forms within five minutes indicating the presence of the polyamine.

2. The method of claim 1, further comprising comparing the intensity of said blue color formed with a predetermined scale for determining the presence or concentration of polyamines in a sample.

3. The method of claim 1, further comprising spectrophotometrically measuring the absorbance of said blue color and said green color and comparing the relative proportion of said blue color and said green color to a working curve obtained by plotting the concentration of said polyamine against relative proportions of said blue color and said green color.

4. The method of claim 1, wherein said polyamine is selected from the group consisting of putrescine, cadaverine, spermidine and spermidine.

5. The method of claim 4, wherein said polyamine is selected from the group consisting of putrescine and cadaverine.

6. The method of claim 1, wherein said aqueous sample is contacted with sodium carbonate to obtain said alkaline pH.

7. The method of claim 6, wherein said sodium carbonate is anhydrous.

8. The method of claim 1, wherein said aqueous sample contains sodium chloride, potassium chloride, lithium chloride or urea.

9. The method of claim 8, wherein said aqueous sample contains sodium chloride.

10. The method of claim 6, wherein the ratio of indigo carmine and sodium carbonate is 1:

1000.

11. The method of claim 3, wherein the absorbance of said blue color is measured at 617 nm and the absorbance of said green color is measured at 778 nm.

12. The method of claim 1, wherein said sample is a vaginal sample from a patient suspected of exhibiting bacterial vaginosis.

13. A test kit for use in a method for the detection of a polyamine associated with bacterial vaginosis, which comprises the following separate components in a container:

(1) a finely ground admixture of indigo carmine and sodium carbonate;

(2) an aqueous solution of sodium chloride, potassium chloride, lithium chloride or urea; and

(3) dimethyl sulfoxide, in amounts sufficient to produce a blue colored solution in the presence of a polyamine.

14. The kit of claim 13, wherein said sodium carbonate is anhydrous.

15. The kit of claim 13, further comprising at least one member selected from the group consisting of (4) a color scale for enabling the presence or concentration of polyamines to be estimated by comparing the blue color formed in the presence of a polyamine with said scale, (5) a sterile swab, and (6) a test vessel adapted for mixing components (1), (2) and (3) in said sample.

16. The kit of claim 13, wherein said aqueous solution is a sodium chloride solution.