Phage-based detection of borreliosis and means therefor

1. A method of determining infection of a subject by either Borrelia burgodorferi sensu lato or Relapsing Fever Borrelia with phage specific for Borrelia by determining presence or the absence of the phage dispensed from and specific to Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia in a blood sample derived from the subject infected or suspected of being infected by Borrelia burgodorferi sensu lato or Relapsing Fever Borrelia , the method comprising the steps of:

a) extracting phage nucleic acid from the blood sample by

i) incubating the Borrelia in ammonium hydroxide and

ii) adding phenol-chloroform to the Borrelia and ammonium hydroxide mixture,

b) detecting the presence or absence of the phage nucleic acid dispensed from and specific to Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia in the blood sample obtained from a subject infected or suspected of being infected by Borrelia burgodorferi sensu lato or Relapsing Fever Borrelia ; and

c) determining the Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia infection of the subject wherein detection of the phage specific to either of Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia in the sample is indicative of infection of the subject of Borrelia burgodorferi sensu lato or Relapsing Fever Borrelia , and the lack of detection of phage dispensed by Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia in the sample indicates the subject is not infected by Borrelia burgodorferi sensu lato or Relapsing Fever Borrelia.

2. The method of claim 1 , wherein the phage nucleic acid encodes a terminase protein.

3. The method of claim 1 , wherein the phage nucleic acid comprises a nucleic acid according to the sequence of:

a) SEQ ID NOS: 1-10; or a nucleic acid with greater than or equal to 70-99.5% sequence homology with SEQ ID NOS: 1-10; or a fragment thereof, or wherein the phage nucleic acid is capable of encoding a protein according to any one of SEQ ID NOS: 36-45 or a fragment thereof, or

b) SEQ ID NOS: 84 or 86; or a nucleic acid with greater than or equal to 70-99.5% sequence homology with SEQ ID NOS: 84 or 86; or a fragment thereof, or wherein the phage nucleic acid is capable of encoding a protein according to any one of SEQ ID NOS: 85 or 87 or a fragment thereof.

4. The method of claim 1 , wherein the phage nucleic acid detected comprises SEQ ID NO. 35; or a nucleic acid with greater than or equal to 70-99.5% sequence homology to SEQ ID NO. 35.

5. The method of claim 1 , wherein the sample is plasma or whole blood.

6. The method of claim 1 , wherein the sample is one that has been obtained in the early or late stage of Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia infection.

7. The method of claim 1 , wherein the Borrelia burgdorferi sensu lato may be any of Borrelia afzelii, Borrelia spielmanii, Borrelia valaisiana, Borrelia garinii, Borrelia finlandensis, Borrelia bugdorferi sensu strictu, Borrelia bissettii, Borrelia bavariensis, Borrelia japonica, Borrelia lusitaniae, Borrelia sinica, Borrelia spielmanii, Borrelia tanukii, Borrelia turdi, Borrelia valaisiana, Borrelia yangtze, Borrelia mayonii, Borrelia carolinensis , and Borrelia andersonii, Borrelia lonestari , and Borrelia Americana or any combination thereof.

8. The method of claim 1 , wherein the Relapsing Fever Borrelia may be any of Borrelia miyamotoi, Borrelia hermsii, Borrelia recurrentis, Borrelia crocidurae, Borrelia duttoni, Borrelia hispanica, Borrelia parkeri and Borrelia turicatae or any combination thereof.

9. The method of claim 1 , wherein the method may determine the presence or the absence of a species ofBorrelia burgdorferi sensu lato or Relapsing Fever Borrelia in the sample, the method comprising the steps of:

a) detecting the presence or absence of the Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia species specific phage in the sample; and

b) determining the presence of the species of the Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia in the sample on the basis of the detection of the species specific phage, or the absence of the species of Borrelia burgdorferi sensu lato or Relapsing Fever Borrelia in the sample on the basis of the lack of detection of the species specific phage.

10. The method of claim 1 , wherein the method additionally comprises treatment of a patient for Lyme disease, the patient's sample having tested positive for Borrelia burgdorferi sensu lato.

11. The method of claim 1 , wherein the method additionally comprises treatment of a patient for Relapsing Fever, the patient's sample having tested positive for Relapsing Fever Borrelia.

12. The method of claim 10 or 11 , wherein treatment comprises administering at least one antibiotic.

13. The method of claim 1 wherein the method further comprises subjecting the isolated nucleic acid to amplification by real time polymerase chain reaction.

14. The method of claim 2 wherein the method further comprises a forward primer which is a nucleic acid comprising SEQ ID NO: 70 to amplify a Borrelia bugdorferi sensu strictu specific terminase gene.

15. The method of claim 14 wherein the method further comprises a reverse primer which is a nucleic acid comprising SEQ ID NO: 71 to amplify the Borrelia bugdorferi sensu strictu specific terminase gene.

16. The method of claim 2 wherein the method further comprises a primer which is a nucleic acid comprising any one of SEQ ID NOS: 88 to 93 to amplify a Borrelia miyamotoi specific terminase gene.

17. The method of claim 16 wherein the method further comprises a forward primer which is a nucleic acid comprising SEQ ID NO: 89 or 92 to amplify the Borrelia miyamotoi specific terminase gene.

18. The method of claim 17 wherein the method further comprises a reverse primer which is a nucleic acid comprising SEQ ID NO: 90 or 93 to amplify the Borrelia miyamotoi specific terminase gene.