FRIDAY, Oct. 14 (HealthDay News) -- Mutant enrichment with 3'-modified oligonucleotides (MEMO) is a highly sensitive technique for detecting cancer mutations, according to a study published online Oct. 5 in The Journal of Molecular Diagnostics.
Seung-Tae Lee, M.D., Ph.D., from the Sungkyunkwan University School of Medicine in Korea, and colleagues described MEMO and evaluated its use for detection of cancer mutations in medical specimens. MEMO'S sensitivity to detect mutations was assessed in genomic DNA extracted from cancer-derived cell lines containing EGFR, BRAF, JAK2, TP53, or KRAS mutations, and a bone marrow sample harboring an NPMI mutation from a patient with acute myeloid leukemia. Primer modification, polymerase chain reaction, sequencing analysis, and other downstream sequencing assays were performed to analyze mutant DNA.
The investigators found that MEMO was a simple and inexpensive enrichment technique based on the use of a 3'-modified oligonucleotide primer. This primer blocked normal allele extension, and enabled mutated allele extension. Downstream Sanger sequencing achieved sensitivities of 10−2 to 10−6, depending on the concentrations and thermodynamics of the primers.
"The potential applications of MEMO include all situations in which minority alleles of clinical significance are present and sensitive detection is required," the lead authors write.
Abstract
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