Abstract
The extraction of virological markers in white blood cells (WBCs) from whole blood—without reagents, electricity, or instruments—is the most important first step for diagnostic testing of infectious diseases in resource-limited settings. Here we develop an integrated microfluidic chip that continuously separates WBCs from whole blood and mechanically ruptures them to extract intracellular proteins and nucleic acids for diagnostic purposes. The integrated chip is assembled with a device that separates WBCs by using differences in blood cell size and a mechanical cell lysis chip with ultra-sharp nanoblade arrays. We demonstrate the performance of the integrated device by quantitatively analyzing the levels of extracted intracellular proteins and genomic DNAs. Our results show that compared with a conventional method, the device yields 120% higher level of total protein amount and similar levels of gDNA (90.3%). To demonstrate its clinical application to human immunodeficiency virus (HIV) diagnostics, the developed chip was used to process blood samples containing HIV-infected cells. Based on PCR results, we demonstrate that the chip can extract HIV proviral DNAs from infected cells with a population as low as 102/μl. These findings suggest that the developed device has potential application in point-of-care testing for infectious diseases in developing countries.
Highlights
Infectious diseases, such as those caused by Human immunodeficiency, Ebola, Hepatitis, Influenza, and Dengue viruses, have been a leading cause of more than 50% of deaths in developing countries over the past decade[1,2,3]
We designed, fabricated, and developed an integrated sample preparation chip made of three polydimethylsiloxane (PDMS) layers and a mechanical cell lysis chip to extract intracellular components of white blood cells (WBCs) from whole blood
Whole blood and phosphate-buffered saline (PBS) buffer are independently injected into the device from the two inlet ports
Summary
Infectious diseases, such as those caused by Human immunodeficiency, Ebola, Hepatitis, Influenza, and Dengue viruses, have been a leading cause of more than 50% of deaths in developing countries over the past decade[1,2,3]. To develop diagnostics that rapidly identify infectious agents to provide timely treatment, the World Health Organization (WHO) has established a set of criteria whose initial letters form the acronym “ASSURED”: (i) affordable, (ii) sensitive, (iii) specific, (iv) user-friendly, (v) rapid and robust, (vi) equipment-free, and (vii) deliverable to those who need them[6]. In response to these demands, various miniaturized diagnostic tools have recently been developed for on-site disease detection. A sample preparation method that is user-friendly, inexpensive, disposable, efficient, and reagent/equipment-free should be developed for use in developing countries
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
