Ekala M T, Lekoulou F, Djikou S, Dubreuil G, Issifou S, Ntoumi F
Centre international de recherches médicales (CIRMF), Gabon.
Bull Soc Pathol Exot. 2000 Feb;93(1):8-11.
In field-based studies, sometimes it is difficult to collect and store samples. We have evaluated a method of malaria parasite deoxyribonucleic (DNA) extraction from non-stained thick dried blood smears collected from 108 Gabonese patients. This method of DNA isolation was compared to those using phenol/chloroform. Patients parasitemia ranged from 0 to 240,000 parasites/microliter of blood. Both methods of DNA preparation gave similar results. Of the 108 slides, 57% were Plasmodium falciparum positive after PCR analysis of the MSA-2 gene and 34% were positive by microscopical examination. Thirty-six and seventy-two blood smears from patients were also tested after one and four weeks' storage respectively, at room temperature, and the parasite DNA was successfully extracted. We conclude that this simple method of collection and rapid procedure of parasite DNA isolation are adequate and convenient in the field when a large number of samples are required and in the case of repetitive samplings of patients.
在实地研究中，有时采集和储存样本很困难。我们评估了一种从108名加蓬患者采集的未染色厚血膜干燥涂片提取疟原虫脱氧核糖核酸（DNA）的方法。将这种DNA分离方法与使用苯酚/氯仿的方法进行了比较。患者的寄生虫血症范围为每微升血液0至240,000个寄生虫。两种DNA制备方法得到了相似的结果。在对MSA-2基因进行PCR分析后，108张载玻片中57%为恶性疟原虫阳性，显微镜检查阳性率为34%。分别在室温下储存1周和4周后，还对36份和72份患者血涂片进行了检测，成功提取了寄生虫DNA。我们得出结论，当需要大量样本以及对患者进行重复采样时，这种简单的采集方法和快速的寄生虫DNA分离程序在实地是足够且方便的。
