Double auxotrophic mycobacterium and uses thereof
Described herein are methods of determining the efficacy of a candidate tuberculosis vaccine and methods of determining the efficacy of a candidate small organic molecule tuberculosis treatment by administering the candidate tuberculosis vaccine or the small organic molecule tuberculosis treatment to an animal which has been infected with an auxotrophic Mycobacterium . The auxotrophic Mycobacterium has a mycobacterial genome in which (i) argB gene, or a gene encoding a mycobacterial ArgB enzyme, has been fully or partially deleted and (ii) metA gene, or a gene encoding a mycobacterial MetA enzyme, has been fully or partially deleted, wherein the Mycobacterium is sterilized in the absence of arginine and methionine). The methods include quantifying the amount of auxotrophic Mycobacterium to determine the efficacy.
1. A method of determining the efficacy of a candidate tuberculosis vaccine, comprising:
administering the candidate tuberculosis vaccine to an animal which has been infected with an auxotrophic Mycobacterium comprising a mycobacterial genome in which (i) argB gene, or a gene encoding a mycobacterial ArgB enzyme, has been fully or partially deleted and (ii) metA gene, or a gene encoding a mycobacterial MetA enzyme, has been fully or partially deleted, wherein the Mycobacterium is sterilized in the absence of arginine and methionine),
quantifying the amount of the auxotrophic Mycobacterium in the animal at one or more time points subsequent to administering the candidate tuberculosis vaccine, and
comparing the amount of auxotrophic Mycobacterium quantified to a predetermined control amount, or to the amount of auxotrophic Mycobacterium infected into the animal, so as to determine the efficacy of the candidate tuberculosis vaccine.
2. The method of claim 1 , wherein the Mycobacterium further comprises a reporter gene insertion in the genome thereof and wherein the amount of auxotrophic Mycobacterium in the animal at one or more time points is quantified by quantifying a reporter signal.
3. A method of determining the efficacy of a candidate small organic molecule tuberculosis treatment, comprising:
administering the small organic molecule tuberculosis treatment to an animal which has been infected with an auxotrophic Mycobacterium comprising a mycobacterial genome in which (i) argB gene, or a gene encoding a mycobacterial ArgB enzyme, has been fully or partially deleted and (ii) metA gene, or a gene encoding a mycobacterial MetA enzyme, has been fully or partially deleted, wherein the Mycobacterium is sterilized in the absence of arginine and methionine),
quantifying the amount of auxotrophic Mycobacterium in the animal at one or more time points subsequent to administering the candidate small organic molecule tuberculosis treatment, and
comparing the amount of auxotrophic Mycobacterium quantified to a predetermined control amount, or to the amount of auxotrophic Mycobacterium infected into the animal, so as to determine the efficacy of the small organic molecule tuberculosis treatment.
4. The method of claim 3 , wherein the Mycobacterium further comprises a reporter gene insertion in the genome thereof and wherein the amount of auxotrophic Mycobacterium in the animal at one or more time points is quantified by quantifying a reporter signal.