Membrane-based, sedimentation-assisted plasma separator for point-of-care applications.
Membrane-based, sedimentation-assisted plasma separator for point-of-care applications.
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DOI：
10.1021/ac402459h
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发表时间：
2013-11-05
影响因子：
7.4
通讯作者：
Bau, Haim H.
中科院分区：
文献类型：
--
作者：
Liu, Changchun;Mauk, Michael;Gross, Robert;Bushman, Frederic D.;Edelstein, Paul H.;Collman, Ronald G.;Bau, Haim H.
通常，高灵敏度、现场护理、临床测试（例如 HIV 病毒载量）需要大量血浆。尽管离心机在临床实验室中广泛用于从全血中分离血浆，但离心通常不适合现场检测。目前还没有合适的替代方法来从毫升血液中分离出相对大量的血浆，而这可能需要满足低丰度目标分子的严格检测限规范。我们报告了一种使用简单、低成本、无泵、基于膜、沉降辅助的血浆分离器，能够在几分钟内从未稀释的全血中分离出相对大量的血浆。该血浆分离器由放置在一次性室中的不对称多孔聚砜膜组成。分离过程利用了血细胞的重力沉降和基于尺寸排阻的过滤。血浆分离器表现出“血液进血浆出”的能力，在不到 7 分钟的时间内从 1.8 mL 未稀释的全血中持续提取 275 ±33.5 μL 血浆。该装置用于从掺有 HIV 病毒的全血中分离载有 HIV 病毒的血浆，对于 35,000、3,500 和 350 拷贝/mL 的病毒载量，回收率分别为 95.5% ± 3.5%、88.0% ± 9.5% 和 81.5% ± 12.1%。分离过程会自动终止，以防止过度溶血。然后将载有HIV的血浆注入我们定制的微流控芯片中进行核酸检测，并成功进行逆转录酶环介导的等温扩增（RT-LAMP），证明血浆足够纯净，可以支持高效核酸扩增。
Often, high sensitivity, point of care, clinical tests, such as HIV viral load, require large volumes of plasma. Although centrifuges are ubiquitously used in clinical laboratories to separate plasma from whole blood, centrifugation is generally inappropriate for on-site testing. Suitable alternatives are not readily available to separate the relatively large volumes of plasma from milliliters of blood that may be needed to meet stringent limit-of-detection specifications for low abundance target molecules. We report on a simple to use, low-cost, pump-free, membrane-based, sedimentation-assisted plasma separator capable of separating a relatively large volume of plasma from undiluted whole blood within minutes. This plasma separator consists of an asymmetric, porous, polysulfone membrane housed in a disposable chamber. The separation process takes advantage of both gravitational sedimentation of blood cells and size exclusion-based filtration. The plasma separator demonstrated a “blood in-plasma out” capability, consistently extracting 275 ±33.5 μL of plasma from 1.8 mL of undiluted whole blood in less than 7 min. The device was used to separate plasma laden with HIV viruses from HIV virus-spiked whole blood with recovery efficiencies of 95.5% ± 3.5%, 88.0% ± 9.5%, and 81.5% ± 12.1% for viral loads of 35,000, 3,500 and 350 copies/mL, respectively. The separation process is self-terminating to prevent excessive hemolysis. The HIV-laden plasma was then injected into our custom-made microfluidic chip for nucleic acid Testing And Was Successfully Subjected To Reverse Transcriptase Loop mediated isothermal amplification (RT-LAMP), demonstrating that the plasma is sufficiently pure to support high efficiency nucleic acid amplification.
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影响因子：
6.2
作者：
Chen, Xing;Cui, Dafu;Chen, Jian
通讯作者：
Chen, Jian
影响因子：
3.1
作者：
Nabatiyan, Arman;Parpia, Zaheer A.;Kelso, David M.
通讯作者：
Kelso, David M.
影响因子：
9.3
作者：
Drosten, Christian;Panning, Marcus;Preiser, Wolfgang
通讯作者：
Preiser, Wolfgang
影响因子：
3.8
作者：
Gross, R;Bilker, WB;Strom, BL
通讯作者：
Strom, BL
影响因子：
6.1
作者：
Dimov, Ivan K.;Basabe-Desmonts, Lourdes;Lee, Luke P.
通讯作者：
Lee, Luke P.