Development of PCR-based test systems for genotyping of single nucleotide polymorphisms in innate immunity and inflammatory response genes
https://doi.org/10.36604/1998-5029-2025-98-109-116
Abstract
Introduction. Identifying single-nucleotide polymorphisms (SNPs) in immune-related genes and examining their associations with specific symptoms, syndromes, and diseases is one of the important directions in medical genetics.
Aim. To develop PCR-based assays for genotyping of selected SNPs in innate immunity and inflammatory response genes (TLR1, TLR4, MBL2, PELI1, IL1B, IL6, IL10, TNF, IFNG).
Materials and methods. SNP selection was based on public databases (PubMed, HapMap, RegulomeDB) and custom-developed software. Primers and probes for high-resolution melting (HRM) analysis of amplicons and for asymmetric Linear-After-The-Exponential (LATE) PCR coupled with molecular beacon probe melting analysis were designed in Vector NTI 11.0. Genomic DNA for genotyping was isolated from peripheral blood of 48 mothers with chronic herpesvirus infections and from umbilical cord blood of their newborns.
Results. Twenty SNPs across the target genes were selected. Oligonucleotide sequences and PCR conditions for genotyping were established.
Conclusions. The developed test systems enable genotyping of selected SNPs in TLR1, TLR4, MBL2, PELI1, IL1B, IL6, IL10, TNF, and IFNG genes, thereby facilitating further association studies with the course and outcomes of pregnancy in women with chronic herpesvirus infections and with developmental abnormalities in their newborns.
About the Authors
O. O. Nekrasova
Far Eastern Scientific Center of Physiology and Pathology of Respiration
Russian Federation
Russian Federation
Olesya O. Nekrasova, PhD (Med.), Senior Staff Scientist, Laboratory of Mechanisms of Virus-Associated Developmental Pathologies
22 Kalinina Str., Blagoveshchensk, 675000
D. A. Gassan
Far Eastern Scientific Center of Physiology and Pathology of Respiration
Russian Federation
Russian Federation
Dina A. Gassan, PhD (Med.), Head of Laboratory, Laboratory of Mechanisms of Virus-Associated Developmental Pathologies
22 Kalinina Str., Blagoveshchensk, 675000
A. V. Konev
Far Eastern Scientific Center of Physiology and Pathology of Respiration
Russian Federation
Russian Federation
Andrey V. Konev, Postgraduate Student, Junior Staff Scientist, Laboratory of Mechanisms of Virus-Associated Developmental Pathologies
22 Kalinina Str., Blagoveshchensk, 675000
K. A. Koneva
Far Eastern Scientific Center of Physiology and Pathology of Respiration
Russian Federation
Russian Federation
Kseniya A. Koneva, Assistant Researcher, Laboratory of Mechanisms of Virus-Associated Developmental Pathologies
22 Kalinina Str., Blagoveshchensk, 675000
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Review
For citations:
Nekrasova O.O., Gassan D.A., Konev A.V., Koneva K.A. Development of PCR-based test systems for genotyping of single nucleotide polymorphisms in innate immunity and inflammatory response genes. Bulletin Physiology and Pathology of Respiration. 2025;(98):109-116. (In Russ.) https://doi.org/10.36604/1998-5029-2025-98-109-116
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