METHODS AND COMPOSITIONS FOR DETECTING PATHOGENIC BACTERIA

• US 20150218614A1
• Filed: 04/17/2015
• Published: 08/06/2015
• Est. Priority Date: 05/18/2012
• Status: Active Grant

• Current Assignee: Washington University In St Louis
• Original Assignee: Washington University In St Louis
• Inventors: Henderson, Jeffrey P., Hung, Chia, Chaturvedi, Kaveri
• Granted Patent: US 9,551,021 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/10 Enterobacteria
• CPC Class Codes: G01N 2333/245 Escherichia (G)
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/56916 Enterobacteria, e.g. shigella, salmonella, klebsiella, serratia

Abstract

The present invention encompasses methods and compositions for detecting pathogenic bacteria. Additionally, the present invention encompasses methods and compositions for catalyzing the dismutation of superoxide radicals. Further, the present invention encompasses methods for determining the antibiotic susceptibility of pathogenic bacteria.

Claims

1. A method for detecting the presence of yersiniabactin-expressing bacteria in a subject, the method comprising the steps of:

a) obtaining a sample from the subject;

b) analyzing the sample, in vitro, for the presence or absence of cupric yersiniabactin, wherein the presence of cupric yersiniabactin is determined by contacting the sample with an antibody specific for cupric yersiniabactin; and

c) identifying the presence of yersiniabactin-expressing bacteria in the subject when cupric yersiniabactin is present in the sample.

2. The method of claim 1, wherein the sample is a urine sample.

3. The method of claim 1, wherein the yersiniabactin-expressing bacteria is uropathogenic bacteria.

4. The method of claim 1, wherein the yersiniabactin-expressing bacteria is uropathogenic E. coli.

5. The method of claim 1, wherein the sample is a urine sample, and the yersiniabactin-expressing bacteria are uropathogenic bacteria.

6. The method of claim 1, wherein the subject has recurrent urinary tract infections.

7. A method for detecting the presence of yersiniabactin-expressing bacteria in a subject, the method comprising the steps of:

a) obtaining a sample from the subject;

b) analyzing the sample, in vitro, for the presence or absence of hydroxphenyl-thiazolinyl-thaizolinyl (HPTT); and

c) identifying the presence of yersiniabactin-expressing bacteria in the subject when HPTT is present in the sample.

8. The method of claim 7, wherein the sample is a urine sample.

9. The method of claim 7, wherein the yersiniabactin-expressing bacteria is uropathogenic bacteria.

10. The method of claim 7, wherein the yersiniabactin-expressing bacteria is uropathogenic E. coli.

11. The method of claim 7, wherein the sample is analyzed using liquid chromatography.

12. The method of claim 7, wherein the presence of HPTT is determined by mass spectrometry.

13. The method of claim 12, wherein the detection of a peak with m/z 307 detects HPTT.

14. The method of claim 7, wherein the presence of HPTT is determined by contacting the sample with an antibody specific for HPTT.

15. The method of claim 7, wherein the subject has recurrent urinary tract infections.

16. A method for determining the antibiotic susceptibility of pathogenic bacteria in a subject, the method comprising:

(a) obtaining a sample for the subject;

(b) analyzing the sample, in vitro, for the presence or absence of siderophores selected from the group comprising yersiniabactin, salmochelin and aerobactin;

(c) identifying the combination of siderophores present wherein the combination indicates susceptibility according to the following;

(i) the presence of yersiniabactin only indicates susceptibility to ciprofloxacin (cipro);

(ii) the presence of yersiniabactin+aerobactin indicates resistance to cipro and trimepthoprim-sulfamethoxazole (TMP-SMZ);

(iii) the presence of yersiniabactin+salmochelin indicates susceptibility to cipro and TMP-SMZ;

(iv) the presence of yersiniabactin+aerobactin+salmochelin indicates susceptibility to cipro;

(v) the presence of aerobactin indicates resistance to cipro; and

(vi) the absence of yersiniabactin+aerobactin+salmochelin indicates susceptibility to cipro.

17. The method of claim 16, wherein the presence or absence of siderophores is determined by detecting one or more nucleic acids involved in the biosynthesis of yersiniabactin, salmochelin and aerobactin.

18. The method of claim 17, wherein yersinibactin is detected by detecting the presence of the nucleic acid fyuA, salmochelin is detected by detecting the presence of the nucleic acid iroN, and aerobactin is detected by detecting the presence of the nucleic acid iucD

19. The method of claim 16, further comprising treating the subject based on the combination of siderophores present.

20. The method of claim 16, wherein the pathogenic bacteria is uropathogenic E. coli.