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在健康人群中使用环介导等温扩增技术对血液和口腔细胞进行基因分型。

Genotyping on blood and buccal cells using loop-mediated isothermal amplification in healthy humans.

作者信息

Drogou Catherine, Sauvet Fabien, Erblang Mégane, Detemmerman Liselot, Derbois Céline, Erkel Marie Claire, Boland Anne, Deleuze Jean François, Gomez-Merino Danielle, Chennaoui Mounir

机构信息

Institut de Recherche Biomédicale des Armées (IRBA), Brétigny sur Orge, France.

Université de Paris, Equipe d'Accueil VIgilance FAtigue et SOmmeil (VIFASOM), EA 7330, Paris, France.

出版信息

Biotechnol Rep (Amst). 2020 May 13;26:e00468. doi: 10.1016/j.btre.2020.e00468. eCollection 2020 Jun.

Abstract

Genetic variations contribute to phenotypic individual vulnerabilities to sleep debt, particularly for five single nucleotide polymorphisms (SNPs). Loop-mediated isothermal amplification and melting curve analysis (LAMP-MC) is a recently developed method to characterize SNPs. The aim of present study was to evaluate the LAMP-MC method on blood and buccal cells for detection of five SNPs of interest in healthy humans. We first analyzed signals obtained from LAMP-MC method on 42 samples. Then we compared the results with those of referent TaqMan method. The LAMP-MC method produced specific melting curves for the five SNPs. A high concordance of genotyping results was observed between the two methods for rs5751876_ADORA2A, rs1800629_TNF-α, rs73598374_ADA and rs228697_PER3 in blood and saliva (Cohen's kappa coefficient >0.80). A good agreement ( = 0.61) was observed for rs4680_COMT in blood only. LAMP-MC is a simple and reliable method to study genetic influences on health, sleep debt-related performance impairments and countermeasures.

摘要

基因变异导致个体对睡眠债的表型易感性存在差异,尤其是对于五个单核苷酸多态性(SNP)而言。环介导等温扩增和熔解曲线分析(LAMP-MC)是一种最近开发的用于鉴定SNP的方法。本研究的目的是评估LAMP-MC方法在血液和颊细胞中检测健康人群中五个感兴趣SNP的效果。我们首先分析了通过LAMP-MC方法从42个样本中获得的信号。然后我们将结果与参照TaqMan方法的结果进行比较。LAMP-MC方法针对这五个SNP产生了特异性熔解曲线。两种方法在血液和唾液中对rs5751876_ADORA2A、rs1800629_TNF-α、rs73598374_ADA和rs228697_PER3的基因分型结果具有高度一致性(科恩kappa系数>0.80)。仅在血液中对rs4680_COMT观察到良好的一致性( = 0.61)。LAMP-MC是一种研究基因对健康、睡眠债相关性能损害及应对措施影响的简单可靠方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00fa/7240324/06ffb359a0f0/gr1.jpg

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