A polymer, microfluidic analyzer for mutation detection

Tae Yoon Lee, Louisiana State University
Kyudong Han, Louisiana State University
Dimitris E. Nikitopoulos, Louisiana State University
Steven S. Soper, Louisiana State University
Mark A. Batzer, Louisiana State University
Michael C. Murphy, Louisiana State University


Recognition of point mutations in codon 12 of the K-ras gene is useful for the early diagnosis of several types of cancer in humans, including colorectal cancer. A microfluidic analyzer, that can detect low-abundance point mutations in human genomic DNA, was developed using a modular approach. Each functional device, including a microfluidic polymerase chain reaction (PCR) reactor, a passive diffusional micromixer reactor, and a microfluidic ligase detection reaction (LDR) reactor, was designed and fabricated in polycarbonate. Interconnects were developed for the passage of analyte between reactors, to ensure adequate spacing for thermal isolation and to facilitate assembly of these reactors using adhesive bonding to form the PCR/LDR processing system. Using the polymer microfluidic system, LDR products were produced from a mixture of mutant DNA to wild type of 1:200 in about 1 h, about half the time needed to execute the same protocol using conventional instrumentation. © 2009 CBMS.