One-step molecular detection of the MYD88 L265P mutation by unlabeled probe genotyping analysis

The aim of our study was to establish an unlabeled probe genotyping approach for rapid detection of the MYD88 L265P mutation in the differential diagnosis of Waldenstrӧm macroglobulinemia patients. Analytical and clinical validation of the assay was performed using serially diluted amplicon-cloned s...

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Published inMolecular and cellular probes Vol. 29; no. 1; pp. 74 - 77
Main Authors Poon, Kok-Siong, Liu, Te-Chih, Tan, Karen Mei-Ling, Koay, Evelyn Siew-Chuan
Format Journal Article
LanguageEnglish
Published England Elsevier Ltd 01.02.2015
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Abstract The aim of our study was to establish an unlabeled probe genotyping approach for rapid detection of the MYD88 L265P mutation in the differential diagnosis of Waldenstrӧm macroglobulinemia patients. Analytical and clinical validation of the assay was performed using serially diluted amplicon-cloned standards, 14 clinical bone marrow aspirate samples, and 30 peripheral blood samples from healthy donors, respectively. The analytical validation results showed that the assay is able to reproducibly identify as low as 10% of the L265P mutant. Clinical validation results showed improved detection sensitivity for the L265P mutation compared to Sanger sequencing. With the simplicity, cost-effectiveness, specificity and rapidity, we foresee that the unlabeled probe HRM assay is a good alternative to substitute current established methods for routine diagnostic testing of the MYD88 L265P mutation.
AbstractList The aim of our study was to establish an unlabeled probe genotyping approach for rapid detection of the MYD88 L265P mutation in the differential diagnosis of Waldenstrӧm macroglobulinemia patients. Analytical and clinical validation of the assay was performed using serially diluted amplicon-cloned standards, 14 clinical bone marrow aspirate samples, and 30 peripheral blood samples from healthy donors, respectively. The analytical validation results showed that the assay is able to reproducibly identify as low as 10% of the L265P mutant. Clinical validation results showed improved detection sensitivity for the L265P mutation compared to Sanger sequencing. With the simplicity, cost-effectiveness, specificity and rapidity, we foresee that the unlabeled probe HRM assay is a good alternative to substitute current established methods for routine diagnostic testing of the MYD88 L265P mutation.
Author Tan, Karen Mei-Ling
Liu, Te-Chih
Poon, Kok-Siong
Koay, Evelyn Siew-Chuan
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  fullname: Poon, Kok-Siong
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  givenname: Te-Chih
  surname: Liu
  fullname: Liu, Te-Chih
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  givenname: Karen Mei-Ling
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  organization: Department of Laboratory Medicine, National University Health System, Singapore
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  givenname: Evelyn Siew-Chuan
  surname: Koay
  fullname: Koay, Evelyn Siew-Chuan
  email: evelyn_koay@nuhs.edu.sg
  organization: Molecular Diagnosis Centre, Department of Laboratory Medicine, National University Health System, Singapore
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Issue 1
Keywords Unlabeled probe
Waldenstrӧm macroglobulinemia
L265P
High-resolution melt
MYD88
Language English
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Snippet The aim of our study was to establish an unlabeled probe genotyping approach for rapid detection of the MYD88 L265P mutation in the differential diagnosis of...
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SubjectTerms Antigens, Differentiation - genetics
Genotype
High-resolution melt
Humans
L265P
Leucine - genetics
Molecular Diagnostic Techniques - economics
Molecular Diagnostic Techniques - methods
Mutation
MYD88
Oligonucleotide Probes - genetics
Proline - genetics
Reproducibility of Results
Sequence Analysis, DNA
Unlabeled probe
Waldenstrom Macroglobulinemia - cerebrospinal fluid
Waldenstrom Macroglobulinemia - diagnosis
Waldenstrom Macroglobulinemia - genetics
Waldenstrӧm macroglobulinemia
Title One-step molecular detection of the MYD88 L265P mutation by unlabeled probe genotyping analysis
URI https://dx.doi.org/10.1016/j.mcp.2014.10.004
https://www.ncbi.nlm.nih.gov/pubmed/25462104
https://search.proquest.com/docview/1655523841
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