التفاصيل البيبلوغرافية
| العنوان: |
CHROMOGENIC MEDIUM FOR THE DETECTION AND IDENTIFICATION OF VANCOMYCIN RESISTANT ENTEROCOCCI AND METHOD THEREFOR |
| Document Number: |
20080145881 |
| تاريخ النشر: |
June 19, 2008 |
| Appl. No: |
11/958827 |
| Application Filed: |
December 18, 2007 |
| مستخلص: |
A microbe-specific medium, containing specific chromogenic substrates, for the detection of vancomycin-resistant enterococci in a biological sample, whereby both the detection and identification of vancomycin-resistant enterococci at the species level is achieved utilizing one sample and one test. |
| Inventors: |
Kircher, Susan (Reisterstown, MD, US); Salomon, Jon E. (Stewartstown, PA, US); Douglas-Mckay, Sheryl (Baltimore, MD, US) |
| Assignees: |
Becton, Dickinson and Company (Franklin Lakes, NJ, US) |
| Claim: |
1. A medium for identifying and distinguishing species of a microorganism placed in the medium comprising: a first substrate conjugated to a first imaging moiety, wherein the first substrate is selected to interact preferentially with a first enzyme that is produced by a first species of microorganism compared to a second enzyme that is produced by a second species of microorganism; and a second substrate conjugated to a second imaging moiety, wherein the second substrate is selected to interact preferentially with the second enzyme that is produced by the second species of microorganism compared to the first enzyme that is produced by the first species of microorganism; wherein the first imaging moiety images a first indicator when the first substrate interacts with the first enzyme and the second imaging moiety images a second indicator when the second substrate interacts with the second enzyme and wherein the first indicator is perceptually different from the second indicator. |
| Claim: |
2. The medium of claim 1, wherein the first imaging moiety is a first chromophore and the second imaging moiety is a second chromophore. |
| Claim: |
3. The medium of claim 2 further comprising a third substrate that interacts with at least a third microorganism to image a third indicator that is perceptually different from either the first indicator or the second indicator. |
| Claim: |
4. The medium of claim 3, wherein the third microorganism produces a normally repressed enzyme in response to the interaction with the third substrate wherein the normally repressed enzyme interacts preferentially with the first substrate to image the first indicator. |
| Claim: |
5. The medium of claim 4 wherein the third microorganism produces an enzyme that is the same or different from the second enzyme and that interacts preferentially with the second substrate to image the second indicator and wherein the first indicator and the second indicator together are the third indicator. |
| Claim: |
6. The medium of claim 1 further comprising inhibitors of non-vancomycin resistant enterococci. |
| Claim: |
7. The medium of claim 6 wherein the inhibitors comprise vancomycin. |
| Claim: |
8. The medium of claim 7 wherein the vancomycin is present in an amount effective to inhibit the growth of at least some non-vancomycin resistant enterococci and not significantly inhibit the growth of vancomycin resistant enterococci. |
| Claim: |
9. The medium of claim 7 further comprising an inhibitor of vanC-containing enterococci wherein the inhibitor of vanC-containing enterococci comprises a cephalosporin. |
| Claim: |
10. The medium of claim 9 wherein the cephalosporin is cefoxitin. |
| Claim: |
11. The medium of claim 10 wherein the cefoxitin is present in an amount effective to suppress the growth of the vanC-containing enterococci. |
| Claim: |
12. The medium of claim 11 wherein the amount of cefoxitin does not significantly suppress the growth of a vancomycin resistant enterococci selected from the group consisting of vancomycin resistant enterococci containing a vanA gene and vancomycin resistant enterococci containing a vanB gene. |
| Claim: |
13. The medium of claim 7 wherein the inhibitors further comprise erythromycin. |
| Claim: |
14. The medium of claim 13, wherein the inhibitors further comprise at least one inhibitor selected from the group consisting of colistin, nalidixic acid, aztreonam, and amphotericin B and combinations thereof. |
| Claim: |
15. The medium of claim 9 wherein the inhibitors further comprise erythromycin. |
| Claim: |
16. The medium of claim 1, wherein the first substrate is alpha-D-glucopyranoside and the first enzyme is alpha-D-glucopyranosidase. |
| Claim: |
17. The medium of claim 16, wherein the first microorganism is a first species of vancomycin resistant enterococci. |
| Claim: |
18. The medium of claim 17 wherein the first species of vancomycin resistant enterococci is E. faecalis. |
| Claim: |
19. The medium of claim 9, wherein the second substrate is beta-D-galactopyranoside and the second enzyme is beta-D-galactopyranosidase. |
| Claim: |
20. The medium of claim 19, wherein the second microorganism is as a second species of vancomycin resistant enterococci. |
| Claim: |
21. The medium of claim 20 wherein the second species of vancomycin resistant enterococci is E. faecium. |
| Claim: |
22. The medium of claim 4, wherein the third substrate is methyl-alpha-D-glucopyranoside. |
| Claim: |
23. The medium of claim 22 wherein the third substrate induces the first enzyme, wherein the first enzyme is alpha-D-glucopyranosidase. |
| Claim: |
24. The medium of claim 23, wherein the at least third microorganism is selected from the group consisting of E. casseliflavus and E. gallinarum. |
| Claim: |
25. A medium for detecting and differentiating species of a microorganisms comprising: a first substrate conjugated to a first imaging moiety, wherein the first substrate preferentially interacts with a first enzyme that is produced by a first species of the microorganism compared to a second enzyme that is produced by a second species of the microorganism; and a second substrate conjugated to a second imaging moiety, wherein the second substrate preferentially interacts with the second enzyme that is produced by the second species of a microorganism compared to the first enzyme that is produced by the first species of the microorganism; an inhibitor of non-vancomycin resistant enterococci comprising vancomycin; and at least one of a third substrate or a cephalosporin, wherein the third substrate induces the production of the first enzyme by at least a third species of the microorganism wherein the first imaging moiety images a first indicator when the first substrate interacts with the first enzyme and the second imaging moiety images a second indicator when the second substrate interacts with the second enzyme, and the combined presence of the first imaging moiety and second imaging moiety image a third indicator in the combined presence of the first enzyme and the second enzyme produced by the at least third species of the microorganism, wherein the first, second and third indicators are perceptually different from each other. |
| Claim: |
26. The medium of claim 25 wherein the first and second imaging moieties are selected from the group consisting of chromophores and fluorophores. |
| Claim: |
27. The medium of claim 26 wherein the first imaging moiety conjugated to the first substrate is a first chromogen and the second imaging moiety conjugated to the second substrate is a second chromogen. |
| Claim: |
28. The medium of claim 25, wherein the first substrate is alpha-D-glucopyranoside and the first enzyme is alpha-D-glucopyranosidase. |
| Claim: |
29. The medium of claim 25, wherein the second substrate is beta-D-galactopyranoside and the second enzyme is beta-D-galactopyranosidase. |
| Claim: |
30. The medium of claim 25, wherein the third substrate is methyl-alpha-D-glucopyranoside. |
| Claim: |
31. The medium of claim 25 wherein the first species of microorganism is a first species of vancomycin resistant enterococci. |
| Claim: |
32. The medium of claim 31 wherein the first species of vancomycin resistant enterococci is E. faecalis. |
| Claim: |
33. The medium of claim 25 wherein the second species of microorganism is a second species of vancomycin resistant enterococci. |
| Claim: |
34. The medium of claim 33 wherein the second species of vancomycin resistant enterococci is E. faecium. |
| Claim: |
35. The medium of claim 25 wherein the at least third species of microorganism is a third species of vancomycin resistant enterococci selected from the group consisting of E. casseliflavus, E. gallinarum and combinations thereof. |
| Claim: |
36. The medium of claim 25 wherein the inhibitor of non-vancomycin resistant enterococci further comprises an inhibitor selected from the group consisting of colistin, nalidixic acid, aztreonam, amphotericin B, and combinations thereof. |
| Claim: |
37. The medium of claim 36 wherein the cephalosporin is cefoxitin. |
| Claim: |
38. The medium of claim 37 wherein the inhibitor of non-vancomycin resistant enterococci further comprise erythromycin. |
| Claim: |
39. A method of detecting the presence of a species of vancomycin resistant enterococci comprising: combining a biological sample with a medium, the medium comprising a first substrate conjugated to a first imaging moiety, wherein the first substrate is selected to preferentially interact with a first enzyme that is produced by a first species of vancomycin resistant enterococci compared to a second enzyme that is produced by a second species of vancomycin resistant enterococci; and a second substrate conjugated to a second imaging moiety, wherein the second substrate preferentially interacts with the second enzyme that is produced by the second species of vancomycin resistant enterococci compared to the first enzyme that is produced by the first species of vancomycin resistant enterococci; at least one inhibitor of non-vancomycin resistant enterococci the at least one inhibitor comprising vancomycin; and at least one of a third substrate or a cephalosporin, wherein the third substrate induces the production of the first enzyme by at least a third species of the vancomycin resistant enterococci wherein the first imaging moiety images a first indicator when the first substrate interacts with the first enzyme and the second imaging moiety images a second indicator when the second substrate interacts with the second enzyme, and the combined presence of the first imaging moiety and second imaging moiety image a third indicator in the combined presence of the first enzyme and the second enzyme produced by the at least third species of the vancomycin resistant enterococci, wherein the first, second and third indicators are perceptually different from each other; and detecting the presence or absence of at least the first and second species of the vancomycin resistant enterococci directly from the medium. |
| Claim: |
40. The medium of claim 39 wherein the first and second imaging moieties are selected from the group consisting of chromophores and fluorophores. |
| Claim: |
41. The method of claim 39 wherein the first imaging moiety conjugated to the first substrate is a first chromogen and the second imaging moiety conjugated to the second substrate is a second chromogen. |
| Claim: |
42. The method of claim 39, wherein the first substrate is alpha-D-glucopyranoside and the first enzyme is alpha-D-glucopyranosidase. |
| Claim: |
43. The method of claim 39, wherein the second substrate is beta-D-galactopyranoside and the second enzyme is beta-D-galactopyranosidase. |
| Claim: |
44. The method of claim 39, wherein the third substrate is methyl-alpha-D-glucopyranoside. |
| Claim: |
45. The method of claim 39 wherein the first species of vancomycin resistant enterococci is E. faecalis. |
| Claim: |
46. The method of claim 39 wherein the second species of vancomycin resistant enterococci is E. faecium. |
| Claim: |
47. The method of claim 39 wherein the third species of vancomycin resistant enterococci is selected from the group consisting of E. casseliflavus, E. gallinarum and combinations thereof. |
| Claim: |
48. The method of claim 39 wherein the inhibitor of non-vancomycin resistant enterococci further comprises an inhibitor selected from the group consisting of colistin, nalidixic acid, aztreonam, erythromycin, amphotericin B, and combinations thereof. |
| Claim: |
49. The method of claim 48 wherein the cephalosporin is cefoxitin. |
| Claim: |
50. The medium of claim 39 wherein the inhibitor of non-vancomycin resistant enterococci further comprises erythromycin. |
| Current U.S. Class: |
435/18 |
| Current International Class: |
12; 12 |
| رقم الانضمام: |
edspap.20080145881 |
| قاعدة البيانات: |
USPTO Patent Applications |