Detection of internal tandem duplications in the FLT3 gene by different electrophoretic methods

Tamás Bubán, Katalin Koczok, Róza Földesi, Gabriella Szabó, Andrea Sümegi, Miklós Tanyi, László Szerafin, Miklós Udvardy, János Kappelmayer, Péter Antal-Szalmás

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: In acute myeloid leukemia (AML), the internal tandem duplication (ITD) in the juxtamembrane domain of the FLT3 (Fms-like tyrosine kinase 3) gene is one of the most frequent genetic alterations associated with poor prognosis. Methods: A complex evaluation of the analytical properties of the three most frequently used detection methods PCR followed by agarose (AGE), polyacrylamide (PAGE) or capillary electrophoresis (CE) was performed on 95 DNA samples obtained from 73 AML patients. Results: All the three methods verified the presence of a mutant allele in 20 samples from 18 patients. AGE and PAGE could detect the presence of 12 mutant allele, while the detection limit of CE was 0.28. However, acceptable reproducibility (inter-assay CV <25) of the mutant allele rate determination was only achievable above 1.5 mutant/total allele rate. The reproducibility of the ITD size determination by CE was much better, but the ITD size calculated by PeakScanner or GeneScan analysis was 7 lower as compared to values obtained by DNA sequencing. The presence of multiple ITD was over-estimated by PAGE and AGE due to the formation of heteroduplexes. Conclusions: This study suggests the use of PCRCE in the diagnostics and the follow-up of AML patients. The data further supports the importance of proper analytical evaluation of home-made molecular biological diagnostic tests.

Original languageEnglish
Pages (from-to)301-310
Number of pages10
JournalClinical Chemistry and Laboratory Medicine
Volume50
Issue number2
DOIs
Publication statusPublished - Feb 1 2012

Keywords

  • acute myeloid leukemia
  • agarose gel electrophoresis
  • capillary electrophoresis
  • fms-like tyrosine kinase 3 (FLT3)
  • internal tandem duplication (ITD)
  • polyacrylamide gel electrophoresis

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Fingerprint Dive into the research topics of 'Detection of internal tandem duplications in the FLT3 gene by different electrophoretic methods'. Together they form a unique fingerprint.

  • Cite this