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比较不同卡片上采集的干血斑中基因组 DNA 的手动提取方法:对临床实践的影响。

Comparison of manual methods of extracting genomic DNA from dried blood spots collected on different cards: implications for clinical practice.

机构信息

Pediatric Clinic 1, Department of Pathophysiology and Transplantation, Universita’ degli Studi di Milano, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy.

出版信息

Int J Immunopathol Pharmacol. 2013 Jul-Sep;26(3):779-83. doi: 10.1177/039463201302600324.

Abstract

Isolating genomic DNA from blood samples is essential when studying the associations between genetic variants and susceptibility to a given clinical condition, or its severity. This study of three extraction techniques and two types of commercially available cards involved 219 children attending our outpatient pediatric clinic for follow-up laboratory tests after they had been hospitalised. An aliquot of venous blood was drawn into plastic tubes without additives and, after several inversions, 80 microL were put on circles of common paper cards and Whatman FTA-treated cards. Three extraction methods were compared: the Qiagen Investigator, Gensolve, and Masterpure. The best method in terms of final DNA yield was Masterpure, which led to a significantly higher yield regardless of the type of card (p less than 0.001), followed by Qiagen Investigator and Gensolve. Masterpure was also the best in terms of price, seemed to be simple and reliable, and required less hands-on time than other techniques. These conclusions support the use of Masterpure in studies that evaluate the associations between genetic variants and the severity or prevalence of infectious diseases.

摘要

从血液样本中提取基因组 DNA 对于研究遗传变异与特定临床疾病易感性或严重程度之间的关联至关重要。本研究涉及三种提取技术和两种市售卡片,共纳入 219 名在我院住院后接受门诊儿科实验室随访检查的儿童。采集静脉血于无添加剂的塑料管中,充分颠倒混匀后,吸取 80μL 于普通滤纸圆片和 Whatman FTA 处理过的滤纸上。比较了三种提取方法:Qiagen Investigator、Gensolve 和 Masterpure。就最终 DNA 产量而言,Masterpure 是最佳方法,无论卡片类型如何,Masterpure 均能显著提高产量(p 均小于 0.001),其次是 Qiagen Investigator 和 Gensolve。Masterpure 在价格方面也具有优势,操作简便可靠,所需时间比其他技术少。这些结论支持在评估遗传变异与传染病严重程度或流行程度之间的关联的研究中使用 Masterpure。

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