Method of rapid analyte detection

• US 5,750,357 A
• Filed: 01/30/1995
• Issued: 05/12/1998
• Est. Priority Date: 05/18/1994
• Status: Expired due to Fees

• Current Assignee: Microquest Diagnostics, Inc.
• Original Assignee: Microquest Diagnostics, Inc.
• Inventors: Olstein, Alan D., Albert, Richard
• Primary Examiner(s): Scheiner, Toni R.
• Application Number:
  US08/380,643
• Time in Patent Office: 1,198 Days
• Field of Search: 435/7.1, 435/7.9, 435/7.32, 435/7.2, 436/501, 436/518, 436/534
• US Class Current: 435/7.32
• 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: G01N 33/56911 Bacteria
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex

Abstract

A detectable synthetic copolymer useful to detect the presence of a microorganism in a test sample is provided. The copolymer comprises repeating monomeric units, which incorporate a population of first monomeric units each comprising a binding agent which binds to a microorganism having multiple binding sites for said binding agent and which further incorporates a population of a second monomeric units each comprising a detectable label or a binding site for a detectable label.

Claims

1. A detectable synthetic copolymer comprising repeating monomeric units, wherein a first population of the monomeric units each comprises a binding agent which binds to a microorganism having multiple binding sites for said binding agent and wherein a second population of the monomeric units each comprises a detectable label or a binding site for a detectable label.

2. The synthetic copolymer of claim 1 wherein the copolymer includes more of said second monomeric units than of said first monomeric units.

3. The synthetic copolymer of claim 1 wherein the microorganism is a pathogenic bacterium.

4. The synthetic copolymer of claim 1 wherein the detectable label or the binding site for the detectable label is bound to each of the population of said second monomeric units via a linking moiety.

5. The synthetic copolymer of claim 2 wherein the detectable label is a fluorescent molecule.

6. The synthetic copolymer of claim 2 wherein the copolymer includes a ten-fold excess of second monomeric units.

7. The synthetic copolymer of claim 5 which comprises a water-insoluble polymeric microsphere.

8. The synthetic copolymer of claim 1 wherein the binding site for the detectable label is biotin.

9. The synthetic copolymer of claim 1 wherein the binding agent is an antibiotic or a fragment thereof.

10. The synthetic copolymer of claim 9 wherein the antibiotic is polymyxin B or the nonapeptide fragment thereof.

20. The synthetic copolymer of claim 1 which is essentially linear.

11. A method for detecting the presence of microorganisms in a test sample comprising:

(a) immobilizing microorganisms from said test sample;

(b) contacting said microorganisms with an amount of a detectable synthetic copolymer which comprises repeating monomeric units, wherein a first population of the monomeric units each comprises a binding agent which binds to said microorganisms at multiple binding site for said binding agent on each microorganism, and wherein a second population of the monomeric units each comprises a detectable label or a binding site for a detectable label, so as to form microorganism-copolymer complexes; and

(c) detecting said microorganism-copolymer complexes.

12. The method of claim 11 wherein the second population of monomeric units is larger than the first population of monomeric units.

13. The method of claim 11 wherein the detectable synthetic copolymer is essentially linear.

14. The method of claim 11 wherein the binding agent is an antibiotic or a fragment thereof, which binds to binding sites on a pathogenic microorganism.

15. The method of claim 14 wherein the microorganism is a bacterium.

16. The method of claim 15 wherein the microorganism is E. coli.

17. The method of claim 14 wherein the detectable label is a fluorogenic molecule.

18. The method of claim 14 wherein the binding site for the detectable label is a biotin molecule.

19. The method of claim 18 wherein the complexes are detected by reaction of the biotin with enzyme labelled avidin followed by a substrate for the enzyme that yields a detectable reaction product.

21. The method of claim 14 wherein the antibiotic is polymyxin B or the nonapeptide fragment thereof.

22. A detectable synthetic copolymer comprising repeating monomeric units prepared by copolymerizing a first population of monomeric units, each comprising a binding agent which binds to a microorganism having multiple binding sites for said binding agent, and a second population of monomeric units, each comprising a detectable label or a binding site for a detectable label.

23. The synthetic copolymer of claim 22 wherein the copolymer includes a greater number of said second monomeric units than of said first monomeric units.

24. The synthetic copolymer of claim 23 wherein the copolymer includes a ten-fold excess of second monomeric units.

25. The synthetic copolymer of claim 22 which is essentially linear.

26. The synthetic copolymer of claim 22 wherein the microorganism is a pathogenic bacterium.

27. The synthetic copolymer of claim 22 wherein the detectable label or the binding site for the detectable label is bound to each of the population of said second monomeric units via a linking moiety.

28. The synthetic copolymer of claim 23 wherein the detectable label is a fluorescent molecule.

29. The synthetic copolymer of claim 28 which comprises a water-insoluble polymeric microsphere.

30. The synthetic copolymer of claim 22 wherein the binding site for the detectable label is biotin.

31. The synthetic copolymer of claim 22 wherein the binding agent is an antibiotic or a fragment thereof.

32. The synthetic copolymer of claim 31 wherein the antibiotic is polymyxin B or the nonapeptide fragment thereof.