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口腔DNA采集:口腔拭子与FTA卡的比较。

Buccal DNA collection: comparison of buccal swabs with FTA cards.

作者信息

Milne Elizabeth, van Bockxmeer Frank M, Robertson Laila, Brisbane Joanna M, Ashton Lesley J, Scott Rodney J, Armstrong Bruce K

机构信息

Telethon Institute for Child Health Research, Centre for Child Health Research, The University of Western Australia.

出版信息

Cancer Epidemiol Biomarkers Prev. 2006 Apr;15(4):816-9. doi: 10.1158/1055-9965.EPI-05-0753.

Abstract

Collection and analysis of DNA, most commonly from blood or buccal cells, is becoming more common in epidemiologic studies. Buccal samples, which are painless to take and relatively easily collected, are often the preferred source. There are several buccal cell collection methods: swabs, brushes, mouthwash, and treated cards, such as FTA or IsoCode cards. Few studies have systematically compared methods of buccal cell collection with respect to DNA yield and amplification success under similar conditions. We compared buccal DNA collection and amplification using buccal swabs and FTA cards in 122 control subjects from our Australian case-control study of childhood acute lymphoblastic leukaemia. Buccal DNA was quantified using a real-time PCR for beta-actin and genotyped at the loci of three polymorphisms (MTHFR 677C>T, ACE I/D, and XPD 1012G>A). PCR was successful with DNA from buccal swabs for 62% to 89% of subjects and from FTA cards for 83% to 100% of subjects, depending on the locus. The matched pair odds ratios (95% confidence interval) comparing success of FTA cards with buccal swabs are as follows: MTHFR 677C>T using PCR-RFLP, 12.5 (11.6-13.5) and using real-time PCR, 130.0 (113.1-152.8); ACE I/D using PCR-amplified fragment length polymorphism, 3.36 (3.2-3.5); XPD 1012G>A using real-time PCR, 150.0 (132.7-172.3). FTA cards are a robust DNA collection method and generally produce DNA suitable for PCR more reliably than buccal swabs. There are, however, technical challenges in handling discs punched from FTA cards that intending users should be aware of.

摘要

在流行病学研究中,最常见的是从血液或颊细胞中收集和分析DNA,这种情况正变得越来越普遍。颊样本采集无痛且相对容易,因此通常是首选来源。有几种颊细胞收集方法:拭子、刷子、漱口水以及经过处理的卡片,如FTA卡或IsoCode卡。很少有研究在相似条件下就DNA产量和扩增成功率对颊细胞收集方法进行系统比较。在我们针对儿童急性淋巴细胞白血病的澳大利亚病例对照研究中,我们对122名对照受试者使用颊拭子和FTA卡进行颊部DNA收集和扩增。使用针对β-肌动蛋白的实时PCR对颊部DNA进行定量,并对三个多态性位点(MTHFR 677C>T、ACE I/D和XPD 1012G>A)进行基因分型。根据位点不同,使用颊拭子采集的DNA,PCR成功的受试者比例为62%至89%;使用FTA卡采集的DNA,PCR成功的受试者比例为83%至100%。比较FTA卡和颊拭子成功率的配对比值比(95%置信区间)如下:使用PCR-RFLP法检测MTHFR 677C>T为12.5(11.6 - 13.5),使用实时PCR法为130.0(113.1 - 152.8);使用PCR扩增片段长度多态性法检测ACE I/D为3.36(3.2 - 3.5);使用实时PCR法检测XPD 1012G>A为150.0(132.7 - 172.3)。FTA卡是一种可靠的DNA收集方法,与颊拭子相比,通常能更可靠地产生适合PCR的DNA。然而,处理从FTA卡上剪下的圆盘存在技术挑战,有意使用的人员应予以注意。

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