Microsatellite-based cancer detection using capillary array electrophoresis and energy-transfer fluorescent primers.
The development of sensitive, rapid, and accurate methods and apparatus for high-throughput short tandem repeat (STR) analysis will be critical for the use of microsatellite alteration in cancer screening. Here we show that STR-based bladder cancer diagnosis can be performed using capillary array electrophoresis and two-color labeling with energy-transfer (ET) fluorescent primers. Rapid (< or = 35 min) separations are achieved on capillary arrays using replaceable separation matrices and the allelic ratios are quantitatively determined with a precision of +/- 10%. With this precision, a variation of 20% was considered diagnostically significant. These methods provide a significant improvement in the speed, ease, and precision of STR analyses compared to slab gel electrophoresis.
Wang
Y
Y
Department of Chemistry, University of California, Berkeley 94720, USA.
Hung
S C
SC
Linn
J F
JF
Steiner
G
G
Glazer
A N
AN
Sidransky
D
D
Mathies
R A
RA
D004273:DNA, Neoplasm / Q000032:analysis*; D004276:DNA, Satellite / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D004735:Energy Transfer; D005456:Fluorescent Dyes; D006801:Humans; D018895:Microsatellite Repeats; D012091:Repetitive Sequences, Nucleic Acid; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis* / Q000235:genetics
D004273:DNA, Neoplasm; D004276:DNA, Satellite; D005456:Fluorescent Dyes