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首页> 外文期刊>Cytokine >Genotyping of single nucleotide polymorphisms in cytokine genes using real-time PCR allelic discrimination technology.
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Genotyping of single nucleotide polymorphisms in cytokine genes using real-time PCR allelic discrimination technology.

机译：使用实时PCR等位基因识别技术对细胞因子基因中的单核苷酸多态性进行基因分型。

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摘要

Single nucleotide polymorphisms (SNPs), particularly those within regulatory regions of genes that code for cytokines often impact expression levels and can be disease modifiers. Investigating associations between cytokine genotype and disease outcome provides valuable insight into disease etiology and potential therapeutic intervention. Traditionally, genotyping for cytokine SNPs has been conducted using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), a low throughput technique not amenable for use in large-scale cytokine SNP association studies. Recently, Taqman((R)) real-time PCR chemistry has been adapted for use in allelic discrimination assays. The present study validated the accuracy and utility of real-time PCR technology for a number of commonly studied cytokine polymorphisms known to influence chronic inflammatory diseases. We show that this technique is amenable to high-throughput genotyping and overcomes many of the problematic features associated with PCR-RFLP including post-PCR manipulation, non-standardized assay conditions, manual allelic identification and false allelic identification due to incomplete enzyme digestion. The real-time PCR assays are highly accurate with an error rate in the present study of <1% and concordance rate with PCR-RFLP genotyping of 99.4%. The public databases of cytokine polymorphisms and validated genotyping assays highlighted in the present study will greatly benefit this important field of research.
机译：单核苷酸多态性（SNP），特别是编码细胞因子的基因调控区域内的单核苷酸多态性，通常会影响表达水平，并且可能成为疾病的修饰因子。研究细胞因子基因型与疾病结局之间的关联为了解疾病病因和潜在的治疗干预提供了宝贵的见识。传统上，已经使用聚合酶链反应限制片段长度多态性（PCR-RFLP）进行了细胞因子SNP的基因分型，这是一种不适用于大规模细胞因子SNP关联研究的低通量技术。近来，实时荧光定量PCR化学已被改编用于等位基因鉴别测定中。本研究验证了实时PCR技术对许多已知会影响慢性炎症疾病的细胞因子多态性的准确性和实用性。我们显示该技术适用于高通量基因分型，并克服了与PCR-RFLP相关的许多问题特征，包括PCR后操作，非标准化测定条件，手动等位基因鉴定和由于不完全酶消化造成的错误等位基因鉴定。实时荧光定量PCR的准确性很高，本研究中的错误率小于1％，PCR-RFLP基因分型的一致性率为99.4％。本研究中重点介绍的细胞因子多态性的公共数据库和经过验证的基因分型分析将极大地有益于这一重要的研究领域。

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