Strøm Gro E A, Tellevik Marit G, Hanevik Kurt, Langeland Nina, Blomberg Bjørn
Department of Clinical Science, University of Bergen, 5020 Bergen, Norway National Centre for Tropical Infectious Diseases, Department of Medicine, Haukeland University Hospital, 5021 Bergen, Norway
Department of Clinical Science, University of Bergen, 5020 Bergen, Norway National Centre for Tropical Infectious Diseases, Department of Medicine, Haukeland University Hospital, 5021 Bergen, Norway.
Trans R Soc Trop Med Hyg. 2014 Aug;108(8):488-94. doi: 10.1093/trstmh/tru084. Epub 2014 Jun 7.
Few studies comparing multiple methods for DNA extraction from dried blood spots (DBS) on filter paper for PCR targeting the Plasmodium genome have been done.
Frequently-used methods for DNA extraction from DBS using Chelex-100, InstaGene Matrix, QIAamp DNA Mini Kit and TE buffer were compared on a dilution series of a standardized Plasmodium falciparum positive sample. The two DNA extraction methods resulting in the lowest limits of detection were compared by testing both on 31 P. falciparum positive samples collected under field conditions and stored for 4 years.
The Chelex-100, InstaGene Matrix and QIAamp DNA Mini Kit methods performed similarly, resulting in the detection of 0.5 to 2 parasites per microliter (p/µl). The same 13 clinical samples (13/31; 42%) were positive using both DNA extraction methods with the lowest limits of detection.
Simple and low-cost methods can be sensitive and useful in extracting DNA from DBS. Poor results on stored clinical DBS indicate that further studies on the impact of storage duration and conditions, and choice of filter paper should be performed.
针对用于靶向疟原虫基因组进行PCR的滤纸干血斑（DBS），比较多种DNA提取方法的研究较少。
在一系列稀释的标准化恶性疟原虫阳性样本上，比较了使用Chelex-100、InstaGene Matrix、QIAamp DNA Mini试剂盒和TE缓冲液从DBS中提取DNA的常用方法。通过对31份在现场条件下采集并保存4年的恶性疟原虫阳性样本进行测试，比较了两种检测限最低的DNA提取方法。
Chelex-100、InstaGene Matrix和QIAamp DNA Mini试剂盒方法表现相似，每微升可检测到0.5至2个疟原虫（p/µl）。两种检测限最低的DNA提取方法对相同的13份临床样本（13/31；42%）检测呈阳性。
简单且低成本的方法在从DBS中提取DNA时可能灵敏且有用。保存的临床DBS样本结果不佳表明，应进一步研究储存时间和条件以及滤纸选择的影响。
