TY - JOUR

T1 - Detection of human genome mutations associated with pregnancy complications using 3-D microarray based on macroporous polymer monoliths

AU - Glotov, A S

AU - Sinitsyna, E S

AU - Danilova, M M

AU - Vashukova, E S

AU - Walter, J G

AU - Stahl, F

AU - Baranov, V S

AU - Vlakh, E G

AU - Tennikova, T B

N1 - Funding information: This work was financially supported by Grant of Russian Scientific Foundation (project 14-50-00069) and by personal Fellowship of President of Russian Federation ( SP-2763.2015.4 ) for Dr. E. Sinitsyna. We also thank Prof. Thomas Scheper for his great help in organization of all measurements and valuable discussions.

PY - 2016/1/15

Y1 - 2016/1/15

N2 - Analysis of variations in DNA structure using a low-density microarray technology for routine diagnostic in evidence-based medicine is still relevant. In this work the applicability of 3-D macroporous monolithic methacrylate-based platforms for detection of different pathogenic genomic substitutions was studied. The detection of nucleotide replacements in F5 (Leiden G/A, rs6025), MTHFR (C/T, rs1801133) and ITGB3 (T/C, rs5918), involved in coagulation, and COMT (C/G, rs4818), TPH2 (T/A, rs11178997), PON1 (T/A rs854560), AGTR2 (C/A, rs11091046) and SERPINE1 (5G/4G, rs1799889), associated with pregnancy complications, was performed. The effect of such parameters as amount and type of oligonucleotide probe, amount of PCR product on signal-to-noise ratio, as well as mismatch discrimination was analyzed. Sensitivity and specificity of mutation detections were coincided and equal to 98.6%. The analysis of SERPINE1 and MTHFR genotypes by both NGS and developed microarray was performed and compared.

AB - Analysis of variations in DNA structure using a low-density microarray technology for routine diagnostic in evidence-based medicine is still relevant. In this work the applicability of 3-D macroporous monolithic methacrylate-based platforms for detection of different pathogenic genomic substitutions was studied. The detection of nucleotide replacements in F5 (Leiden G/A, rs6025), MTHFR (C/T, rs1801133) and ITGB3 (T/C, rs5918), involved in coagulation, and COMT (C/G, rs4818), TPH2 (T/A, rs11178997), PON1 (T/A rs854560), AGTR2 (C/A, rs11091046) and SERPINE1 (5G/4G, rs1799889), associated with pregnancy complications, was performed. The effect of such parameters as amount and type of oligonucleotide probe, amount of PCR product on signal-to-noise ratio, as well as mismatch discrimination was analyzed. Sensitivity and specificity of mutation detections were coincided and equal to 98.6%. The analysis of SERPINE1 and MTHFR genotypes by both NGS and developed microarray was performed and compared.

KW - Aryldialkylphosphatase/genetics

KW - Base Sequence

KW - Catechol O-Methyltransferase/genetics

KW - Ethylene Glycols

KW - Female

KW - Genome, Human

KW - Genotype

KW - Humans

KW - Integrin beta3/genetics

KW - Methacrylates/chemistry

KW - Methylenetetrahydrofolate Reductase (NADPH2)/genetics

KW - Mutation

KW - Oligonucleotide Array Sequence Analysis

KW - Plasminogen Activator Inhibitor 1/genetics

KW - Porosity

KW - Pregnancy

KW - Pregnancy Complications/genetics

KW - Tryptophan Hydroxylase/genetics

KW - Mutations

KW - Macroporous polymer monoliths

KW - Pregnancy complications

KW - DNA microarrays

KW - Single nucleotide polymorphism (SNP)

UR - http://www.scopus.com/inward/record.url?scp=84945431364&partnerID=8YFLogxK

U2 - 10.1016/j.talanta.2015.09.066

DO - 10.1016/j.talanta.2015.09.066

M3 - Article

C2 - 26592644

VL - 147

SP - 537

EP - 546

JO - TALANTA

JF - TALANTA

SN - 0039-9140

ER -