Membrane-Based, Sedimentation-Assisted Plasma Separator for Point-of-Care Applications
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- 作者：Changchun Liu ; Michael Mauk ; Robert Gross ; Frederic D. Bushman ; Paul H. Edelstein ; Ronald G. Collman ; Haim H. Bau
- 刊名：Analytical Chemistry
- 出版年：2013
- 出版时间：November 5, 2013
- 年：2013
- 卷：85
- 期：21
- 页码：10463-10470
- 全文大小：376K
- 年卷期：v.85,no.21(November 5, 2013)
- ISSN：1520-6882
文摘
Often, high-sensitivity, point-of-care (POC) 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 鈥渂lood in鈥損lasma out鈥?capability, consistently extracting 275 卤 33.5 渭L of plasma from 1.8 mL of undiluted whole blood within 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% 卤 3.5%, 88.0% 卤 9.5%, and 81.5% 卤 12.1% for viral loads of 35鈥?00, 3500, 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.