Colorimetric approach to high-throughput mutation analysis.

High-throughput genomic mutation screening for primary tumors has characteristically been expensive, labor-intensive, and inadequate to detect low levels of mutation in a background of wild-type signal. We present a new, combined PCR and colorimetric approach that is inexpensive, simple, and can detect the presence of 1% mutation in a background of wild-type. We compared manual dideoxy sequencing of p53 for eight lung cancer samples to a novel assay combining a primer extension step and an enzymatic colorimetric step in a 96-well plate with covalently attached oligonucleotide sequences.

A new technology for mutation detection.

A sensitive, accurate, and simple method, called shifted termination assay (STA), was developed for detection of genetic mutations. The STA technology can be used to detect genetic mutations in polymerase chain reaction (PCR)-amplified samples of tissue, and plasma and serum that include circulating DNA containing point mutations, insertions, deletions, translocations, or single nucleotide polymorphisms (SNPs). STA is a multiple-base and multiple-cycle primer extension based detection method that can identify mutant DNA in samples containing as little as 1% mutant DNA in a mixture with 99% wild-type DNA.