The invention provides a method of detecting surrogate markers for
active tuberculosis in a serum sample. The surrogate markers are selected from serum mycolic acid
antigen, serum anti-mycolic acid antibodies or both. The method includes the steps of combining the serum sample with a labelled
monoclonal immunoglobulin
antibody or fragment thereof to mycolic acids to produce a combined serum sample, the
antibody or fragment thereof not substantially cross-reacting with
cholesterol and the
label being selected so that binding of the labelled
antibody to immobilized mycolic acid
antigen of mycobacterial origin produces a detectable
signal and combining a blank sample with the labelled
monoclonal immunoglobulin antibody or fragment thereof to mycolic acid to produce a combined blank sample. The method includes exposing both samples to immobilised mycolic acid
antigen of mycobacterial origin or a
synthetic analogue or analogues thereof so that the labelled immunoglobulin antibodies or fragments thereof in each sample bind to the immobilised antigen to produce detectable signals. If the surrogate markers are present, the
signal produced by the blank sample will be stronger than that produced by the serum sample because of inhibition of binding of the labelled antibody in the serum sample arising from prior binding of the labelled antibody with the mycolic acid antigen in the serum sample or by
competitive binding of serum anti-mycolic acid antibodies in the serum sample to the immobilised mycolic acid antigen or both.