We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Detection of the MYD88 mutation by the combination of the allele-specific PCR and quenching probe methods.
- Authors
Nogami, S.; Kawaguchi‐Ihara, N.; Shiratori, E.; Ohtaka, M.; Itoh, M.; Tohda, S.
- Abstract
Introduction The MYD88 missense mutation c.794T>C, p.Leu265Pro, is found in patients with Waldenstörm's macroglobulinemia and lymphoma. Direct sequencing, allele-specific PCR ( AS- PCR), PCR-restriction fragment length polymorphism ( PCR- RFLP), and high-resolution melting analysis ( HRM) are currently used to detect the mutation; however, they are either time-consuming or have low detection sensitivity. Here, we developed a novel highly sensitive and rapid detection method based on the quenching probe ( QP) technique and AS- PCR. Method A lymphoma cell line heterozygous for the MYD88 mutation, two wild-type cell lines, and two samples from Waldenstörm's macroglobulinemia patients were analyzed by AS- PCR, PCR- RFLP, HRM, and QP, and their detection sensitivity was examined using the mixtures of the mutant and wild-type DNA. Results For mutation-carrying heterozygous samples, the QP method produced W-shaped melting profiles presenting curves derived from the wild-type and mutant alleles. The QP analysis was performed in 2 h and demonstrated the detection limit of 5%, which was similar to that of the other methods. However, the combination of AS- PCR and QP ( AS- QP) improved the sensitivity to 0.62% of the mutant allele. Conclusion The AS- QP analysis is rapid and minimally improves detection sensitivity compared to the AS- PCR.
- Subjects
ALLELES; CELL physiology; DNA fingerprinting; GENES; GENETICS; LYMPHOMAS; GENETIC mutation; POLYMERASE chain reaction; PROTEINS; RESEARCH funding; GENETIC carriers; DATA analysis software; GENOTYPES
- Publication
International Journal of Laboratory Hematology, 2017, Vol 39, Issue 2, p163
- ISSN
1751-5521
- Publication type
Article
- DOI
10.1111/ijlh.12598