acl-based detection of non-A. fumigatus species. (A) PCR amplification and gel electrophoresis of various acl genes among non-A. fumigatus species. The species (from top to bottom) and assays are shown as follows: A. flavus (amplification of the 403-bp aclFl1-3′ region; A. flavus-specific amplicons are marked with arrowheads, and an amplicon detected in A. parasiticus is marked with an asterisk); A. nidulans (amplification of the 227-bp aclN1-3′ region); A. niger (amplification of the 297-bp aclNi1-5′ region); and A. terreus (amplification of the 262-bp aclT2-5′ conserved region). PCR amplification was resolved by 2% agarose gel electrophoresis. M, 1-kb Plus TrackIt DNA ladder. (B) Nanogel electrophoretic separation. PCR amplicons specific for A. fumigatus, A. flavus, A. nidulans, A. niger, and A. terreus were simultaneously resolved and detected with the intercalating dye SYBR green 1. The separation is accomplished using a 25-μm-inner-diameter capillary with an effective length of 40.2 cm, E = 100 V/cm, 30°C, and a 10% nanogel with [DMPC]/[DHPC] = 2.5 at 6 kV for a 2-s injection. RFU, relative fluorescence units.