Rapid and highly sensitive detection of malaria-infected erythrocytes using a cell microarray chip.
Rapid and highly sensitive detection of malaria-infected erythrocytes using a cell microarray chip.
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DOI：
10.1371/journal.pone.0013179
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发表时间：
2010-10-13
期刊：
影响因子：
3.7
通讯作者：
Kataoka M
中科院分区：
文献类型：
--
作者：
Yatsushiro S;Yamamura S;Yamaguchi Y;Shinohara Y;Tamiya E;Horii T;Baba Y;Kataoka M
疟疾是许多流行国家的主要人类传染病之一。为了防止疟疾的传播，有必要开发一种早期、灵敏、准确和常规的诊断系统。细胞微阵列芯片用于检测感染疟疾的红细胞。该芯片由聚苯乙烯制成，有20944个微腔(宽105微米，深50微米)，观察到微腔中红细胞单层的形成。以培养的恶性疟原虫3D7株为研究对象，验证了该细胞芯片在疟疾诊断中的应用潜力。将核染色染料Syto 59中的红细胞悬浮液分散在芯片表面，然后静置10分钟，使红细胞稳定进入微腔。每个微室容纳约130个红细胞，在一个芯片上总共有超过270万个红细胞。用微阵列扫描仪在5分钟内检测到任何荧光阳性的红细胞，可检测到0.0001%的寄生虫血症。为检测人血中纯化红细胞对白细胞的污染情况，将20微克L全血与10毫升RPMI1640混合，通过白细胞分离滤器。洗脱部分以1,000×g离心2min，小球分散在1.0ml的培养液中。将Syto 59加入红细胞悬浮液中，然后在细胞微阵列芯片上进行分析。观察到细胞在微腔中也有类似的适应。在细胞芯片上，污染的白细胞数小于1个。结果表明，该芯片可用于疟疾感染红细胞的检测，其灵敏度是常规光学显微镜的10-100倍，操作简便，纯化红细胞只需15min即可完成。
Malaria is one of the major human infectious diseases in many endemic countries. For prevention of the spread of malaria, it is necessary to develop an early, sensitive, accurate and conventional diagnosis system. A cell microarray chip was used to detect for malaria-infected erythrocytes. The chip, with 20,944 microchambers (105 µm width and 50 µm depth), was made from polystyrene, and the formation of monolayers of erythrocytes in the microchambers was observed. Cultured Plasmodium falciparum strain 3D7 was used to examine the potential of the cell microarray chip for malaria diagnosis. An erythrocyte suspension in a nuclear staining dye, SYTO 59, was dispersed on the chip surface, followed by 10 min standing to allow the erythrocytes to settle down into the microchambers. About 130 erythrocytes were accommodated in each microchamber, there being over 2,700,000 erythrocytes in total on a chip. A microarray scanner was employed to detect any fluorescence-positive erythrocytes within 5 min, and 0.0001% parasitemia could be detected. To examine the contamination by leukocytes of purified erythrocytes from human blood, 20 µl of whole blood was mixed with 10 ml of RPMI 1640, and the mixture was passed through a leukocyte isolation filter. The eluted portion was centrifuged at 1,000×g for 2 min, and the pellet was dispersed in 1.0 ml of medium. SYTO 59 was added to the erythrocyte suspension, followed by analysis on a cell microarray chip. Similar accommodation of cells in the microchambers was observed. The number of contaminating leukocytes was less than 1 on a cell microarray chip. The potential of the cell microarray chip for the detection of malaria-infected erythrocytes was shown, it offering 10–100 times higher sensitivity than that of conventional light microscopy and easy operation in 15 min with purified erythrocytes.
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影响因子：
3
作者：
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通讯作者：
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影响因子：
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作者：
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DOI：
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发表时间：
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期刊：
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影响因子：
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