Alternately binding probe competitive PCR as a simple, cost-effective, and accurate quantification method for JAK2V617F allele burden in myeloproliferative neoplasms

Leuk Res. 2011 Dec;35(12):1632-6. doi: 10.1016/j.leukres.2011.06.016. Epub 2011 Jul 12.

Abstract

We developed a simple, cost-effective, and accurate JAK2 allele burden quantification method named alternately binding probe competitive PCR (ABC-PCR). ABC-PCR can be performed to quantify target JAK2 allele burdens in a single reaction. The throughput and running cost of ABC-PCR are markedly improved compared with those of allele-specific quantitative PCR (AS-qPCR). The quantification of samples with known JAK2 allele burdens revealed that ABC-PCR had a small assay-to-assay variation. The JAK2 allele burdens in the patients with myeloproliferative neoplasms measured by ABC-PCR and AS-qPCR showed a good fitting. ABC-PCR would be a powerful tool for quantifying target JAK2 allele burdens.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Alleles
  • Amino Acid Substitution / genetics
  • Binding, Competitive
  • Bone Marrow Neoplasms / genetics*
  • Cell Line, Tumor
  • Cost-Benefit Analysis
  • DNA Mutational Analysis / methods
  • DNA Mutational Analysis / standards
  • DNA Probes / chemistry
  • DNA Probes / metabolism
  • Gene Dosage
  • Humans
  • Janus Kinase 2 / analysis
  • Janus Kinase 2 / genetics*
  • Models, Biological
  • Myeloproliferative Disorders / genetics
  • Phenylalanine / genetics
  • Polymerase Chain Reaction / economics*
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards
  • Reference Standards
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Valine / genetics

Substances

  • DNA Probes
  • Phenylalanine
  • JAK2 protein, human
  • Janus Kinase 2
  • Valine