Beta lactamase as biomarker for the specific detection of tuberculosis-complex bacteria
The present disclosure provides methods, reagents, systems, and devices that target β lactamase as a biomarker for the sensitive and specific detection of tuberculosis-complex bacteria. Specifically, the present disclosure relates to methods and compositions for the detection of specific β-lactamase protein and nucleic acid sequences to indicate the presence of tuberculosis-complex bacteria.
1. A method of detecting and treating the presence of tuberculosis-complex bacterial infection in a subject, the method comprising:
determining the presence or amount of anti-tuberculosis-complex β-lactamase (BlaC) antibody in a biological sample obtained from the subject, wherein the presence or amount of anti-BlaC antibody in the biological sample is indicative of the presence of tuberculosis-complex bacterial infection in the subject, and
treating the subject indicated to have a tuberculosis-complex bacterial infection.
2. The method of claim 1 , wherein the presence of anti-tuberculosis-complex BlaC antibody in the biological sample is determined by an assay comprising the steps:
(a) contacting the biological sample with at least one polypeptide with an amino acid sequence that has at least 90% sequence identity to any six or more contiguous amino acids of SEQ ID NO:2; and
(b) detecting the formation of a complex between the antibody in the sample and the polypeptide.
3. The method of claim 1 , further comprising comparing the determined amount of anti-tuberculosis-complex BlaC antibody to a reference standard, where an amount of anti-tuberculosis-complex BlaC antibody detected in the biological sample greater than the reference standard is indicative of the presence of tuberculosis-complex bacteria in the subject.
4. The method of claim 3 , wherein the reference standard is an analogous biological sample from a subject that does not have tuberculosis-complex bacteria.
5. The method of claim 1 , wherein the tuberculosis-complex bacteria are from one or more of the species selected from the group consisting of: Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium bovis - Bacillus Calmette-Guerin (BCG), Mycobacterium africanum, Mycobacterium microti, Mycobacterium canettii, Mycobacterium pinnipedii , and Mycobacterium mungi.
6. The method of claim 1 , wherein the subject is a mammal.
7. The method of claim 1 , wherein the subject is human.
8. The method of claim 1 , wherein the biological sample is selected from the group consisting of blood, serum, sputum, saliva, breath, feces, urine, spinal fluid, mucus or tissue sample.
9. The method of claim 2 , wherein the polypeptide is attached to a substrate.
10. The method of claim 9 , wherein the substrate is a particle, a filter, or a well.
11. The method of claim 9 , wherein the attachment is through a linker.
12. The method of claim 2 , wherein the polypeptide is detectably labeled.
13. The method of claim 3 , wherein the reference standard is determined from a biological sample obtained from the subject at or prior to the commencement of treatment for a tuberculosis infection.
14. The method of claim 2 , wherein the at least one polypeptide has an amino acid sequence that has at least 95% sequence identity to any six or more contiguous amino acids of SEQ ID NO:2.
15. The method of claim 2 , wherein the at least one polypeptide has an amino acid sequence that has at least 95% sequence identity to any 15 or more contiguous amino acids of SEQ ID NO:2.
16. The method of claim 2 , wherein the at least one polypeptide comprises amino acids corresponding to amino acids at positions 75 to 87 of SEQ ID NO:2, at amino acids at positions 142 to 144 of SEQ ID NO:2, and/or at amino acids at positions 249 to 252 of SEQ ID NO:2.