Jiangsu Institute of Parasitic Diseases, Meiyuan Yangxiang 117, Wuxi 214064, People's Republic of China.
Malar J. 2011 Aug 25;10:251. doi: 10.1186/1475-2875-10-251.
Many malaria-related studies depend on infected red blood cells (iRBCs) as fundamental material; however, infected blood samples from human or animal models include leukocytes (white blood cells or WBCs), especially difficult to separate from iRBCs in cases involving Plasmodium vivax. These host WBCs are a source of contamination in biology, immunology and molecular biology studies, requiring their removal. Non-woven fabric (NWF) has the ability to adsorb leukocytes and is already used as filtration material to deplete WBCs for blood transfusion and surgery. The present study describes the development and evaluation of a prototype NWF filter designed for purifying iRBCs from malaria-infected blood.
Blood samples of P. vivax patients were processed separately by NWF filter and CF11 column methods. WBCs and RBCs were counted, parasite density, morphology and developing stage was checked by microscopy, and compared before and after treatment. The viability of filtrated P. vivax parasites was examined by in vitro short-term cultivation.
A total of 15 P. vivax-infected blood samples were treated by both NWF filter and CF11 methods. The WBC removal rate of the NWF filter method was 99.03%, significantly higher than the CF11 methods (98.41%, P < 0.01). The RBC recovery rate of the NWF filter method was 95.48%, also significantly higher than the CF11 method (87.05%, P < 0.01). Fourteen in vitro short-term culture results showed that after filter treatment, P. vivax parasite could develop as normal as CF11 method, and no obvious density, developing stage difference were fund between two methods.
NWF filter filtration removed most leukocytes from malaria-infected blood, and the recovery rate of RBCs was higher than with CF11 column method. Filtrated P. vivax parasites were morphologically normal, viable, and suitable for short-term in vitro culture. NWF filter filtration is simple, fast and robust, and is ideal for purification of malaria-infected blood.
许多疟疾相关的研究都依赖于感染的红细胞（iRBC）作为基础材料；然而，来自人类或动物模型的感染血液样本包括白细胞（白细胞或 WBC），特别是在涉及间日疟原虫的情况下，很难与 iRBC 分离。这些宿主白细胞是生物学、免疫学和分子生物学研究中的污染来源，需要去除。无纺织物（NWF）具有吸附白细胞的能力，已经用作过滤材料，以去除用于输血和手术的白细胞。本研究描述了一种用于从感染疟疾的血液中纯化 iRBC 的原型 NWF 过滤器的开发和评估。
分别用 NWF 过滤器和 CF11 柱法处理间日疟原虫患者的血液样本。通过显微镜检查计数白细胞和红细胞，检查寄生虫密度、形态和发育阶段，并在治疗前后进行比较。通过体外短期培养检查过滤后的间日疟原虫的活力。
共处理了 15 份间日疟原虫感染的血液样本，分别用 NWF 过滤器和 CF11 方法处理。NWF 过滤器方法的白细胞去除率为 99.03%，明显高于 CF11 方法（98.41%，P<0.01）。NWF 过滤器方法的红细胞回收率为 95.48%，也明显高于 CF11 方法（87.05%，P<0.01）。14 个体外短期培养结果表明，过滤处理后，间日疟原虫可以像 CF11 方法一样正常发育，两种方法之间没有明显的密度和发育阶段差异。
NWF 过滤器过滤从感染疟疾的血液中去除了大部分白细胞，红细胞的回收率高于 CF11 柱法。过滤后的间日疟原虫形态正常，具有活力，适合短期体外培养。NWF 过滤器过滤简单、快速、稳健，是纯化感染疟疾的血液的理想方法。
