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Journal of Biomedical Research | Volume: 29
November 19-20, 2018 | Paris, France
Molecular Biology, Tissue Science and Regenerative Medicine
International Conference on
Joint Event
&
4
th
World Heart Congress
Diagnostic biochips for the analysis of tumor biomarkers
T V Nasedkina
Engelhardt Institute of Molecular Biology, Russia
G
enetic aberrations in leukemia often lead to the formation
of expressed chimeric genes, which should be assessed
for proper diagnosis and therapy. We developed a biochip-
based assay for the analysis of 22 most clinically relevant
fusion genes occurring in pediatric leukemia, including 8
recurrent translocations involved the MLL gene and different
gene partners. The method includes a multiplex reverse
transcriptase–polymerase chain reaction (RT–PCR) to amplify
and fluorescently label a fusion transcript fragments and
subsequent hybridization of on a biochip with immobilized
oligonucleotides complementary to different parts of fusion
genes. The fine structure of the MLL fusion genes, including
localization of the MLL breakpoints, was performed in 43
infants and 28 pediatric cases. Other important cancer
biomarkers predicting prognosis and therapeutic targets are
somatic mutations in solid tumors. A diagnostic biochip for
the detection of 39 clinically relevant somatic mutations in the
BRAF, NRAS, KIT, GNAQ, GNA11, MAP2K1 and MAP2K2 genes
has been developed. The multiplex LNA-clamp PCR was used
for the preferable amplification of mutated over wild type DNA.
The approach could detect 0.5% of mutated DNA in the sample
analyzed.
The biochip-based assay is a robust and highly sensitivemethod
for the detection of fusion genes and somatic mutations in
cancer patients.
e:
nased@biochip.ruMolecular Biology & Heart Congress 2018, Volume 29
DOI: 10.4066/biomedicalresearch-C8-023