Detection of IL28B SNP DNA from buccal epithelial cells, small amounts of serum, and dried blood spots.
Detection of IL28B SNP DNA from buccal epithelial cells, small amounts of serum, and dried blood spots.
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DOI：
10.1371/journal.pone.0033000
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Bourlière M
中科院分区：
文献类型：
--
作者：
Halfon P;Ouzan D;Khiri H;Pénaranda G;Castellani P;Oulès V;Kahloun A;Amrani N;Fanteria L;Martineau A;Naldi L;Bourlière M
白细胞介素28基因（rs 12979860）编码区的点突变最近已被确定用于预测丙型肝炎病毒感染的治疗结果。该多态性检测基于全血DNA提取。或者，用于基因诊断的DNA来源于颊上皮细胞（BEC）、干血斑（DBS）和来自血清的基因组DNA。本研究的目的是调查单核苷酸多态性等位基因rs 12979860 CC的替代检测途径的可靠性和准确性。提取200例患者的血液、血浆和血清样本（400 µL）。使用FTA卡测试口腔涂片。为了模拟邮政延迟，我们测试了在环境温度下储存对五个时间点（基线、48小时、6天、9天和12天）的不同来源的DNA的影响。血液、血浆、血清和BEC之间的一致性为100%，验证了从细胞学刷上收集的BEC提取的DNA用于基因检测的用途。应用涂片技术检测血涂片（3620拷贝）和口腔涂片（5870拷贝）中HPTR 1基因的遗传变异。这些结果与以1/10稀释的全血的结果相似。分别需要至少0.04 µL、4 µL和40 µL才能获得全血、血清和血浆的可利用结果。对于不同来源的DNA，在每个时间点之间未观察到显著变化。在这些不同时间点的IL 28 B SNP分析显示使用四种DNA来源的相同结果。我们证明，从口腔细胞、少量血清和干血斑中提取基因组DNA是从外周血细胞中提取DNA的替代方法，有助于对多种遗传标记进行回顾性和前瞻性研究，特别是在难以接触的个体中。
Point mutations in the coding region of the interleukin 28 gene (rs12979860) have recently been identified for predicting the outcome of treatment of hepatitis C virus infection. This polymorphism detection was based on whole blood DNA extraction. Alternatively, DNA for genetic diagnosis has been derived from buccal epithelial cells (BEC), dried blood spots (DBS), and genomic DNA from serum. The aim of the study was to investigate the reliability and accuracy of alternative routes of testing for single nucleotide polymorphism allele rs12979860CC. Blood, plasma, and sera samples from 200 patients were extracted (400 µL). Buccal smears were tested using an FTA card. To simulate postal delay, we tested the influence of storage at ambient temperature on the different sources of DNA at five time points (baseline, 48 h, 6 days, 9 days, and 12 days) There was 100% concordance between blood, plasma, sera, and BEC, validating the use of DNA extracted from BEC collected on cytology brushes for genetic testing. Genetic variations in HPTR1 gene were detected using smear technique in blood smear (3620 copies) as well as in buccal smears (5870 copies). These results are similar to those for whole blood diluted at 1/10. A minimum of 0.04 µL, 4 µL, and 40 µL was necessary to obtain exploitable results respectively for whole blood, sera, and plasma. No significant variation between each time point was observed for the different sources of DNA. IL28B SNPs analysis at these different time points showed the same results using the four sources of DNA. We demonstrated that genomic DNA extraction from buccal cells, small amounts of serum, and dried blood spots is an alternative to DNA extracted from peripheral blood cells and is helpful in retrospective and prospective studies for multiple genetic markers, specifically in hard-to-reach individuals.
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影响因子：
29.4
作者：
McCarthy JJ;Li JH;Thompson A;Suchindran S;Lao XQ;Patel K;Tillmann HL;Muir AJ;McHutchison JG
通讯作者：
McHutchison JG
影响因子：
3.7
作者：
de Jonge HJ;Fehrmann RS;de Bont ES;Hofstra RM;Gerbens F;Kamps WA;de Vries EG;van der Zee AG;te Meerman GJ;ter Elst A
通讯作者：
ter Elst A
DOI：
10.1097/meg.0b013e328349d0ef
发表时间：
2011-10-01
影响因子：
2.1
作者：
Halfon, Philippe;Bourliere, Marc;Dessein, Alain
通讯作者：
Dessein, Alain
影响因子：
1.8
作者：
McClure, Matthew C;McKay, Stephanie D;Taylor, Jeremy F
通讯作者：
Taylor, Jeremy F
影响因子：
29.4
作者：
Rauch, Andri;Kutalik, Zoltan;Bochud, Pierre-Yves
通讯作者：
Bochud, Pierre-Yves