Nucleic acid purification from dried blood spot on FTA Elute Card provides template for polymerase chain reaction for highly sensitive Plasmodium detection
Nucleic acid purification from dried blood spot on FTA Elute Card provides template for polymerase chain reaction for highly sensitive Plasmodium detection
复制标题
DOI：
10.1016/j.parint.2019.101941
复制
发表时间：
2019-12-01
影响因子：
1.9
通讯作者：
Kataoka, Masatoshi
中科院分区：
文献类型：
--
作者：
Hashimoto, Muneaki;Bando, Mika;Kataoka, Masatoshi
聚合酶链式反应(PCR)是高度敏感地检测疟疾患者感染疟原虫红细胞的重要诊断方法。本研究比较了用于制备干血斑(DBS)的滤纸在聚合酶链式反应检测疟原虫中的性能。在全血中加入恶性疟原虫感染的红细胞，以获取不同程度寄生虫血症的样本，将其应用于Whatman 3 mm Chr试纸、FTA卡片或FTA Elute卡片，以制备DBS。用DNA纯化试剂盒从DBS中提取DNA作为套式聚合酶链式反应的模板。在概率分析中，Whatman 3 mm CHR试纸和FTA卡片的估计检出限(LOD)为5.5只寄生虫/亩L血，而FTA洗脱卡的估计检出限为1.6只/亩L血。这一结果表明，在FTA Elute Card上制备的DBS产生了用于随后基于高灵敏度的PCR检测恶性疟原虫感染的红细胞的最佳模板DNA。这一发现有助于提高疟疾诊断测试的准确性。
Polymerase chain reaction (PCR) is an essential diagnostic method for highly sensitive detection of Plasmodium infected erythrocytes in patients with malaria. This study compared the performance of filter papers used for the preparation of dried blood spots (DBS) in detecting Plasmodium by PCR. Whole blood spiked with P. falciparum-infected erythrocytes to obtain samples with various levels of parasitemia were applied to Whatman 3MM Chr papers, FTA Cards, or FTA Elute Cards to prepare the DBS. DNA was purified from the DBS using a DNA purification kit and used as the template for nested PCR. In probit analysis, the estimated limit of detection (LoD) was 5.5 parasites/mu L blood for Whatman 3MM Chr papers and FTA Cards and 1.6 parasites/mu L blood for the FTA Elute Card. This result suggested that the DBS prepared on an FTA Elute Card yield the best template DNA for subsequent high-sensitivity PCR-based detection of P. falciparum-infected erythrocytes. This finding can help improve the accuracy of malarial diagnostic tests.