The 1L of MUCF, containing mycobacterial proteins of interest, was concentrated at 4 [degrees] C under [N.sub.2] by using a stirred-cell apparatus (Amicon Inc., Beverly, MA) with a 10-kDa molecular weight membrane.
Aliquots of 8 g of MUCF protein were resolved by discontinuous sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) (10) by using 10% to 20% gradient 10-well gels (Novex, San Diego, CA).
The N-terminal amino acid sequences of MUCF proteins were determined and compared with known mycobacterial sequences.
These skin-tested BU patient and control populations from the Daloa region were then tested for reactivity of their sera to MUCF by Western blot, and Burulin induration was compared with serum antibody response (Table 2).
The antibody response to the MUCF was then determined for all 61 clinically diagnosed BU patients and 27 healthy controls, and antibody reactivity to individual antigens was recorded.
We found that a specific response to the MUCF was observed in BU patient sera, but unlike the Burulin DTH response, the antibody response did not correlate with disease stage (Table 2).