Department of Psychiatry, All India Institute of Medical Sciences, New Delhi, India.
Indian J Med Res. 2012 Mar;135(3):318-21.
BACKGROUND & OBJECTIVES: Dried blood spotted on to filter paper has been found suitable for a large number of studies. In tropical countries with varying temperature conditions the use of dried blood needs to be validated. We carried out this study to assess the use of blood spotted filter paper as a transport system to study genotyping of Apo E gene.
Fifty five patients visiting Cardiothoracic Neuroscience Centre (CNC) OPD at the All India Institute of Medical Sciences (AIIMS), New Delhi, and referred for lipid investigations to Cardiac Biochemistry Laboratory were selected at random. Blood was spotted on to Whatman 3 MM filter paper, dried and stored at room temperature. Genomic DNA was extracted and genotyping was carried out at the end of 0, 3 and 12 months. The study was further validated using samples collected on to filter paper from four centres and stored for eight years at room temperature. The temperature and humidity conditions of the centre varied widely.
Fifty five samples collected on to filter paper showed exact match of the genotyping when compared to fresh blood. In dried blood samples collected and stored for 1 yr at room temperature DNA extraction and apo E genotyping was done successfully.
INTERPRETATION & CONCLUSIONS: The present results showed the feasibility of using dried blood samples on filter paper for apo E genotyping in tropical temperature. The findings need to be validated on a large sample before being recommended for use.
在滤纸上采集的干血斑已被证明适用于大量研究。在温度条件变化较大的热带国家,需要对干血的使用进行验证。我们进行这项研究是为了评估将血液斑滤纸用作研究载体制备载脂蛋白 E 基因基因分型的适用性。
我们随机选择了 55 名在新德里全印医学科学院(AIIMS)胸心神经科门诊就诊并因脂质检查而被转至心脏生物化学实验室的患者。将血液点在沃特曼 3MM 滤纸上,干燥后在室温下储存。在 0、3 和 12 个月时提取基因组 DNA 并进行基因分型。使用在室温下储存 8 年的来自四个中心的滤纸上采集的样本进一步验证了该研究。中心的温度和湿度条件差异很大。
与新鲜血液相比,55 个滤纸上采集的干血斑样本的基因分型完全匹配。在室温下收集并储存 1 年的干血样本中,成功进行了 DNA 提取和载脂蛋白 E 基因分型。
目前的结果表明,在热带温度条件下,使用滤纸上的干血斑进行载脂蛋白 E 基因分型是可行的。在推荐使用之前,需要在更大的样本上验证这些发现。