TY - JOUR

T1 - Silica-coated iron oxide nanoparticles for DNA isolation for molecular genetic studies in hematology

AU - Тюменцева, Анна Владимировна

AU - Ярославцев, Роман

AU - Столяр, Сергей Викторович

AU - Саитова, Алина Тимуровна

AU - Тютрина, Екатерина

AU - Горбенко, Алексей

AU - Столяр, Марина

AU - Ольховский, Игорь

N1 - Publisher Copyright: © Copyright 2021, Mary Ann Liebert, Inc., publishers 2021.

PY - 2021/9/17

Y1 - 2021/9/17

N2 - Aim: To develop magnetic nanoparticles (MNPs) based on iron oxide for DNA isolation from blood cells for quantitative molecular genetic analyses of the V617F mutation in the Januskinase 2 (JAK2) gene. Materials and Methods: MNPs were synthesized by the coprecipitation method and coated with tetraethyl orthosilicate (TEOS). The size and shape of the complexes were estimated using transmission electron microscopy. Twenty blood samples from patients with myeloproliferative disorders were used for DNA isolation with the MNPs. DNA quality and compatibility for molecular genetic studies of the JAK2 V617F mutation were investigated by gel electrophoresis and real-time polymerase chain reaction (RT-PCR). Results: The average amount of DNA isolated from 150 μL of whole blood was 75.2 ng when MNPs were used and 72.5 ng when standard silica sorbent was used. There was no DNA damage observed after interaction with MNPs. RT-PCR demonstrated similar values for the JAK2 V617F mutant DNA ratios in the samples after DNA isolation with MNPs and by standard sorption on silica. Conclusions: MNPs with silicate capsules of sufficient thickness were obtained and the undesirable damaging effect of iron oxides on nucleic acids during isolation from cells were eliminated. Designed MNPs allow obtaining intact DNA for molecular genetic studies using the example of the JAK2 V617F for study.

AB - Aim: To develop magnetic nanoparticles (MNPs) based on iron oxide for DNA isolation from blood cells for quantitative molecular genetic analyses of the V617F mutation in the Januskinase 2 (JAK2) gene. Materials and Methods: MNPs were synthesized by the coprecipitation method and coated with tetraethyl orthosilicate (TEOS). The size and shape of the complexes were estimated using transmission electron microscopy. Twenty blood samples from patients with myeloproliferative disorders were used for DNA isolation with the MNPs. DNA quality and compatibility for molecular genetic studies of the JAK2 V617F mutation were investigated by gel electrophoresis and real-time polymerase chain reaction (RT-PCR). Results: The average amount of DNA isolated from 150 μL of whole blood was 75.2 ng when MNPs were used and 72.5 ng when standard silica sorbent was used. There was no DNA damage observed after interaction with MNPs. RT-PCR demonstrated similar values for the JAK2 V617F mutant DNA ratios in the samples after DNA isolation with MNPs and by standard sorption on silica. Conclusions: MNPs with silicate capsules of sufficient thickness were obtained and the undesirable damaging effect of iron oxides on nucleic acids during isolation from cells were eliminated. Designed MNPs allow obtaining intact DNA for molecular genetic studies using the example of the JAK2 V617F for study.

KW - DNA isolation

KW - JAK2 V617F

KW - magnetic nanoparticles

KW - myeloproliferative neoplasms

KW - OXIDATIVE STRESS

KW - MUTATION

KW - DAMAGE

UR - http://www.scopus.com/inward/record.url?scp=85116057116&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/e431249a-f738-3e5a-a554-132ef0c16794/

U2 - 10.1089/gtmb.2021.0067

DO - 10.1089/gtmb.2021.0067

M3 - Article

VL - 25

SP - 611

EP - 614

JO - Genetic Testing and Molecular Biomarkers

JF - Genetic Testing and Molecular Biomarkers

SN - 1945-0265

IS - 9

ER -