量油尺
恶性疟原虫
寄生虫血症
聚合酶链反应
色谱法
洗脱
核酸
生物
间日疟原虫
滤纸
分子生物学
全血
干血
病毒学
疟疾
化学
生物化学
免疫学
尿
基因
作者
Muneaki Hashimoto,Mika Bando,Jun‐ichi Kido,Kazumichi Yokota,Toshihiro Mita,Kazuaki Kajimoto,Masatoshi Kataoka
标识
DOI:10.1016/j.parint.2019.101941
摘要
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/μL blood for Whatman 3MM Chr papers and FTA Cards and 1.6 parasites/μL 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.
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