Composition and method for stabilizing and maintaining the viability of hardy microorganisms

1. A method for preserving viable hardy bacteria in a biological sample at ambient temperature, comprising contacting the biological sample with a stabilization composition to form a mixture, and storing the mixture at ambient temperature, wherein the stabilization composition comprises a chelating agent, a denaturing agent, an inorganic salt and has a pH between 6 and 11,

wherein

the hardy bacteria are pathogenic bacteria comprising one or more species of Mycobacterium and/or Bacillus anthracis,

the denaturing agent is an anionic detergent or a non-ionic detergent,

the inorganic salt comprises lithium chloride, lithium bromide, lithium iodide, lithium acetate, or any combination thereof,

the chelating agent is ethylene glycol tetraacetic acid (EGTA), (2-Hydroxyethyl)ethylenediaminetriacetic acid (HEDTA), diethylene triamine pentaacetic acid (DTPA), nitrilotriacetic acid (NTA), ethylenediaminetriacetic acid (EDTA), cyclohexanediaminetetraacetic acid (CDTA), N,N-bis(carboxymethyl)glycine, citrate anhydrous, sodium citrate, calcium citrate, ammonium citrate, ammonium bicitrate, citric acid, diammonium citrate, ferric ammonium citrate, lithium citrate, or any combination thereof; and

the stabilizing composition does not comprise a reducing agent,

wherein storing the mixture at ambient temperature comprises storing the mixture at a temperature within the range of from 4° C. to 40° C.,

wherein the stabilization composition preserves, in a viable state, and hardy bacteria in the biological sample and reduces or eliminates other microbial growth in the sample.

2. The method of claim 1 , wherein the one or more Mycobacterium is Mycobacterium tuberculosis and wherein the Bacillus anthracis is stabilized as spores.

3. The method of claim 2 , wherein the Mycobacteria hardy bacteria are Mycobacterium tuberculosis.

4. The method of claim 1 , wherein the biological sample is a mucoid bodily fluid.

5. The method of claim 4 , wherein the mucoid biological fluid is sputum or saliva.

6. The method of claim 1 , wherein the composition comprises (i) 2% SDS, 12.5 mM CDTA, 250 mM LiCl, 50 mM glycine and has pH 10.5; or (ii) 4% SDS, 50 mM CDTA, 250 mM LiCl, 140 mM LiOH and has pH 6.8.

7. The method of claim 1 , wherein all or a portion of the hardy bacteria remain stable following storage at room temperature for 1 day or more, 2 days or more, 3 days or more, 4 days or more, 5 days or more, 6 days or more or 7 days or 1 month or more.

8. The method according to claim 1 , wherein the stabilizing composition liquefies the biological sample.

9. The method of claim 8 , wherein the denaturing agent is an anionic detergent.

10. The method of claim 9 , wherein the anionic detergent is sodium dodecyl sulfate (SDS), lithium dodecyl sulphate or sodium lauroyl sulfate (SLS).

11. The method of claim 8 , wherein the denaturing agent is a non-ionic detergent.

12. The method of claim 8 , wherein the biological sample is a mucoid bodily fluid.

13. The method of claim 12 , wherein the mucoid bodily fluid is sputum or saliva.

14. The method of claim 8 , wherein the composition comprises (i) 2% SDS, 12.5 mM CDTA, 250 mM LiCl, 50 mM glycine and has pH 10.5; or (ii) 4% SDS, 50 mM CDTA, 250 mM LiCl, 140 mM LiOH and has pH 6.8.

15. The method of claim 1 , wherein the denaturing agent is an anionic detergent.

16. The method of claim 15 , wherein the anionic detergent is sodium dodecyl sulfate (SDS), lithium dodecyl sulphate or sodium lauroyl sulfate (SLS).

17. The method of claim 1 , wherein the denaturing agent is a non-ionic detergent.