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Identification of oncogene mutations in leukemia patients using microchip-based PCR analysis

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Abstract

At present, cytogenetic and PCR-based techniques are generally used in the diagnosis of leukemia. Due to high accuracy, specificity, and sensitivity of PCR assays, they have the advantage over traditional cytogenetic tools. As a rule, the classical real time PCR is carried out in a 25-μL reaction mixture. It requires a large volume of each reagent and takes a long time to finish the test. The molecular genetic assay presented here is microchip-based real-time PCR that is optimized for simultaneous analysis of 15 oncogene mutations and one housekeeping gene abl. Moreover, this diagnostic tool requires a minimal amount of cDNA and PCR reagents (10-fold less) and a short time (2-fold less) for quantitative estimation of more than five copies of gene-target. Thus, microchip-based PCR analysis can improve the detection of oncogene mutations in leukemia patients and may be used for both early diagnostics and long-term monitoring of leukemia.

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Abbreviations

MCT:

molecular colonies technique

MRD:

minimal residual disease

RT:

reverse transcription

ALL:

acute lymphoblastic leukemia

B-ALL:

B-cell ALL

AML:

acute myeloid leukemia

APML:

acute promyelocytic leukemia

CML:

chronic myelogenous leukemia

FISH:

fluorescent in situ hybridization

MALDI-TOF:

time-of-flight matrix-assisted laser desorption/ionization mass spectrometry

NGS:

next generation (whole-genome) sequencing.

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Authors and Affiliations

  1. Institute of Hematology, Almazov Federal Medical Research Center, St. Petersburg, 197341, Russia

    K. V. Bogdanov, T. S. Nikulina, E. G. Lomaia & A. Y. Zaritskey

  2. Lumex Group, St. Petersburg, 192029, Russia

    K. V. Bogdanov & M. N. Slyadnev

Authors
  1. K. V. Bogdanov
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  2. T. S. Nikulina
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  3. E. G. Lomaia
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  4. M. N. Slyadnev
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  5. A. Y. Zaritskey
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Corresponding author

Correspondence to K. V. Bogdanov.

Additional information

Original Russian Text © K.V. Bogdanov, T.S. Nikulina, E.G. Lomaia, M.N. Slyadnev, A.Y. Zaritskey, 2017, published in Bioorganicheskaya Khimiya, 2017, Vol. 43, No. 5, pp. 523–531.

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Bogdanov, K.V., Nikulina, T.S., Lomaia, E.G. et al. Identification of oncogene mutations in leukemia patients using microchip-based PCR analysis. Russ J Bioorg Chem 43, 544–551 (2017). https://doi.org/10.1134/S1068162017040033

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  • DOI: https://doi.org/10.1134/S1068162017040033

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