Compositions and Methods for Culturing Spirochetes

Abstract

The present invention relates to methods for culturing spirochetes, in particular Borrelia burgdorferi . The present invention also provides methods of identifying spirochetes present in a biological sample. The present invention further provides methods of diagnosing diseases cause by a spirochete infection, such as Lyme disease, syphilis, and multiple sclerosis. The present invention further provides methods for identifying spirochete susceptibilities to antimicrobials and antimicrobial compositions and cocktails. The present invention also provides methods for treating subjects suspected of having a spirochete infection.

1 - 30 . (canceled) 
     
     
         31 . A composition for culturing spirochetes comprising a spirochete-supporting medium, serum, a reducing agent, and an antibiotic. 
     
     
         32 . An in vitro method of culturing a spirochete comprising:
 (1) inoculating a composition according to claim  1  with the spirochete to generate a short-term culture; and   (2) incubating the short-term culture in a short-term culture vessel for about 4-6 days.   
     
     
         33 . A method of identifying a spirochete in a subject comprising:
 (1) culturing the spirochete by the method of claim  2 ; and   (2) identifying the spirochete in an identification assay.   
     
     
         34 . A method of diagnosing a spirochete infection in a subject comprising:
 (1) culturing the spirochete by the method according to claim  2 , wherein the spirochete is obtained from a sample from a subject possibly suffering from a spirochete infection; and   (2) detecting the spirochete in a detection assay,   wherein detection of the spirochete indicates that the subject is suffering from a spirochete infection.   
     
     
         35 . A method of identifying the antibiotic sensitivity of a spirochete comprising:
 (1) inoculating a spirochete culture medium in a culture vessel with a spirochete, wherein the spirochete culture medium is a composition according to claim  1 ;   (2) incubating the inoculated medium for about 4-6 days;   (3) transferring the cultured spirochetes into a secondary culture comprising a test antibiotic; and   (4) incubating the secondary culture,   wherein inhibition of the growth or proliferation, or even death, of the spirochete indicates that the spirochete is sensitive to the test antibiotic.   
     
     
         36 . A method of treating a subject suffering from a spirochete infection comprising
 (1) obtaining a sample from a subject suffering from a spirochete infection;   (2) inoculating a spirochete culture medium in a culture vessel with the sample, wherein the spirochete culture medium is a composition according to claims  1 ;   (3) incubating the inoculated medium for about 4-6 days;   (4) transferring the cultured spirochetes into a secondary culture comprising a test antibiotic;   (5) incubating the secondary culture, wherein inhibition of the growth or proliferation, or even death, of the spirochete indicates that the spirochete is sensitive to the test antibiotic; and   (6) administering an effective amount of the test antibiotic to the subject.