METHOD AND KIT FOR DETECTION OF MYCOBACTERIA

20170242005

15/503647

August 13, 2015

A method of determining whether an individual is infected with mycobacteria. The method comprising the steps of (a) providing a system which comprises at least two different mycolic-acid derived antigens; (b) introducing a sample obtained from the individual into the system and into contact with each of the at least two different mycolic-acid derived antigens; and (c) detecting the presence or absence of the binding of a biomarker in the sample with each antigen in the system. The method may be particularly suitable for determining the presence or absence of a disease antibody indicative of infection with any disease caused by infection with mycobacteria, for example tuberculosis, leprosy, pulmonary disease, burili ulcer, Johne's disease and bovine tuberculosis. A kit and device are also described. The present invention may provide an over the counter device to allow individuals to check on a routine basis that they have normal immune responses.

1. A method of determining whether an individual is infected with mycobacteria, the method comprising: (a) providing a system which comprises at least two different mycolic-acid derived antigens; (b) introducing a sample obtained from the individual into the system and into contact with each of the at least two different mycolic-acid derived antigens; and (c) detecting the presence or absence of the binding of a biomarker in the sample with each antigen in the system.

2. The method according to claim 1, wherein the system comprises a substrate which carries the at least two different mycolic-acid derived antigens at different positions on the substrate.

3. The method according to claim 2, wherein the method comprises the steps of: (a) providing a substrate which carries at least two different mycolic-acid derived antigens at different positions; (b) contacting the substrate with a sample obtained from the individual; and (c) detecting the presence or absence of the binding of a biomarker in the sample with each antigen at each position on the substrate.

4. The method according to claim 1, wherein the biomarker is an antibody indicative of infection with a mycobacterial disease.

5. The method according to claim 1, wherein each of the two or more antigens is selected from one or more of the following classes of compounds: (i) mycolic acids obtained from natural sources; (ii) synthetically prepared mycolic acids; (iii) salts of mycolic acids; (iv) esters of mycolic acids (i) and/or (ii); (v) sulfur-containing mycolic acid compounds and/or salts or esters thereof; (vi) simple structural analogues of mycolic acids and/or salts or esters thereof; and (vii) mycolic acid wax esters and/or salts or esters thereof.

6. The method according to claim 1 wherein each antigen is at least 90% pure.

7. The method according to claim 1 wherein the system comprises from 6 to 12 different mycolic-acid derived antigens.

8. The method according to claim 1 wherein the system comprises: (a) a trehalose monomycolate antigen; and/or (b) a trehalose dimycolate antigen; and/or (c) a mycolic acid wax ester antigen; and/or (d) a glucose monomycolate antigen.

9. The method according to claim 6, wherein the system comprises (i) an arabinose mycolate antigen.

10. The method according to claim 1 wherein the system comprises a substrate which carries the at least two different mycolic-acid derived antigens at different positions on the substrate; wherein step (c) involves the steps: (i) contacting the substrate with a composition comprising a secondary antibody; and (ii) observing the substrate at the two or more different positions.

11. The method according to claim 1 which determines the likelihood that an individual is infected with one or more species of mycobacteria.

12. The method according to claim 1 which is a method of determining whether an individual is infected with Mycobacterium avium paratuberculosis; and wherein at least one of the mycolic acid derived antigens is a wax ester or a derivative thereof.

13. A kit for determining the presence or absence of a biomarker in a sample, the kit comprising: (x) a system which comprises two or more different mycolic-acid derived antigens; and (y) a composition comprising a secondary antibody.

14. A device comprising a housing and a system; wherein the system comprises at least two mycolic-acid derived antigens and is located within the housing.

15. The device according to claim 14, wherein the system comprises from 6 to 12 different mycolic-acid derived antigens, the different mycolic-acid derived antigens being selected from: (i) mycolic acids and their esters obtained from natural sources; (ii) synthetically prepared mycolic acids; (iii) salts of mycolic acids; (iv) esters of mycolic acids (i) and/or (ii); (v) sulfur-containing mycolic acid compounds and/or salts or esters thereof; (vi) simple structural analogues of mycolic acids and/or salts or esters thereof; and (vii) mycolic acid wax esters and/or salts or esters thereof.

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edspap.20170242005