Molecular Biology

, Volume 52, Issue 2, pp 206–211 | Cite as

Multiplex Genotyping of Allelic Variants of Genes Involved in Metabolizing Antileukemic Drugs

  • D. O. Fesenko
  • M. A. Avdonina
  • L. G. Gukasyan
  • S. A. Surzhikov
  • A. V. Chudinov
  • A. S. Zasedatelev
  • T. V. Nasedkina
Genomics. Transcriptomics

Abstract

A biochip, primer set, and genotyping protocol were developed to simultaneously address 16 single nucleotide polymorphisms in antileukemic drug metabolism genes, including TPMT, ITPA, MTHFR, SLCO1B1, SLC19A1, NR3C1, GRIA1, ASNS, MTRR, and ABCB1. The genotyping procedure included a one-round multiplex polymerase chain reaction (PCR) with simultaneous incorporation of a fluorescent label into the PCR product and subsequent hybridization on a biochip with immobilized probes. The method was used to test 65 DNA samples of leukemia patients. Fluorescence signal intensity ratios in pairs of wildtype and respective mutant sequence probes were analyzed for all polymorphic markers and demonstrated high accuracy of genotyping. The reliability of genotype determination using the biochip was confirmed by direct Sanger sequencing.

Keywords

genotyping pharmacogenetics biochip biotransformation genes single nucleotide polymorphism single-round PCR 

Abbreviations

MTX

methotrexate

AL

acute leukemia

ALL

acute lymphoblastic leukemia

PCR

polymerase chain reaction

ABCB1

ATP binding cassette subfamily B member 1 gene

ASNS

asparagine synthetase (glutamine-hydrolyzing) gene

dNTP

deoxynucleoside triphosphate

dUTP

deoxyuridine triphosphate

GRIA1

glutamate ionotropic receptor AMPA type subunit 1 gene

ITPA

inosine triphosphate pyrophosphatase gene

SLCO1B1

solute carrier organic anion transporter family member 1B1 gene

MTHFR

methylenetetrahydrofolate reductase gene

MTRR

5-methyltetrahydrofolate–homocysteine methyltransferase reductase gene

NR3C1

nuclear receptor subfamily 3 group C member 1 gene

SLC19A1

solute carrier family 19 member 1 gene

TPMT

thiopurine S-methyltransferase gene

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Copyright information

© Pleiades Publishing, Inc. 2018

Authors and Affiliations

  • D. O. Fesenko
    • 1
  • M. A. Avdonina
    • 1
  • L. G. Gukasyan
    • 1
  • S. A. Surzhikov
    • 1
  • A. V. Chudinov
    • 1
  • A. S. Zasedatelev
    • 1
  • T. V. Nasedkina
    • 1
  1. 1.Engelhardt Institute of Molecular BiologyRussian Academy of SciencesMoscowRussia

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