Provided herein is an assay for detecting a rough-type mycobacterium and a smooth-type mycobacterium in a sample, wherein the mycobacterium is Mycobacterium tuberculosis or Mycobacterium bovis. The assay comprises a first molecule that selectively binds to the rough-type mycobacterium or binds to a molecule preferentially secreted by the rough-type mycobacterium and a second molecule that selectively binds to the smooth-type mycobacterium or binds to a molecule preferentially secreted by the smooth-type mycobacterium. The first molecule and the second molecule are independently detectable. Also provided herein are corresponding methods for detecting a rough-type mycobacterium and a smooth-type mycobacterium in a sample and assays and methods for detecting tuberculosis infection in a sample.
