Awards

Ayokunle Olanrewaju, and collaborators Ashleigh Theberge and Erwin Berthier, of the University of Washington in the U.S. will develop a platform for at-home self-collection of blood, serum separation, and sample stabilization at sufficient sample volumes for comprehensive HIV monitoring. An existing device for home blood collection will be expanded with the development of serum separation using a simple filtration system and connected to a standard blood collection tube with serum-stabilizing reagents. The device design will be optimized to ensure that over 1 mL of blood can be processed. The resulting design will then be tested for its effectiveness for RNA and protein analysis to monitor HIV viral load and biomarkers associated with HIV treatment and care. Performance of the device will be compared to standard blood processing, using blood from healthy volunteers spiked with either HIV RNA or C-reactive protein as a model biomarker. They envision a system that can readily integrate with standard laboratory or point-of-care diagnostic workflows to enable maximal deployability.

Jianghong Rao of Stanford University in the U.S. will develop a magnet-based system for capturing and concentrating the TB bacterium from sputum biosamples to facilitate TB diagnosis. The system is based on conjugating magnetic nanoparticles to bacteriophage specific to Mycobacterium tuberculosis, so that a simple magnet can capture phage-bound TB bacteria from patient sputum. This process will be optimized, including the stability and activity of the magnetized phage in a lyophilized powder form and at a high ambient temperature. A prototype device for the magnetic phage system will be designed and tested for its ability to generate purified target bacteria ready for lysis and PCR-based diagnostic testing. Initial tests will use a non-TB Mycobacterium species, and subsequent tests in collaboration with Niaz Banaei at Stanford Health Care Clinical Microbiology Laboratory will use clinical TB patient samples.

Suman Chakraborty of the Indian Institute of Technology Kharagpur in India will develop a membrane filtration system for Mycobacterium tuberculosis lysis and DNA purification from patient samples to enhance TB diagnosis. A paper-based membrane will be impregnated with two chemical agents, each in a separate layer, for sequential sample processing. Cell lysis will be performed in the first layer by zinc oxide nanoflowers, nanoscale structures that lyse bacterial cells through both mechanical and chemical mechanisms. DNA purification will be performed in the second layer by silica nanoparticles. This membrane system can be attached to a DNA amplification chamber with lyophilized reagents for colorimetric Loop-Mediated Isothermal Amplification (LAMP). This instrument-free, integrated process for TB diagnosis will be tested in a research setting and a clinical pathology laboratory setting.

David Erickson of Cornell University in the U.S. will develop a device for heat inactivation and mechanical lysis of Mycobacterium tuberculosis from patient samples to enhance TB diagnosis. A prototype device will be engineered, using a non-TB Mycobacterium for testing as a proxy. The device combines an alternating magnetic field and magnetic beads to inductively heat inactivate bacterial samples and actuate lysis by a bead-beating mechanism. The protype will be pilot tested in a collaboration at the Infectious Diseases Institute in Kampala, Uganda where patient samples with presumptive TB will be processed with the standard protocol for TB diagnosis and in parallel with the prototype.

Sam Nugen of Cornell University in the U.S. will develop a bacteriophage-based system for the rapid concentration and lysis of Mycobacterium tuberculosis from patient samples to enhance TB diagnosis. A mycobacteriophage will be engineered to express the streptavidin protein, enabling low-cost magnetic particles to capture and concentrate the phage along with the TB bacterium to which it naturally binds. The phage will also be engineered to accelerate lysis of the TB bacterium after it is bound, and phage replication genes will be deleted to ensure that the phage can only replicate in a modified host bacterium or an in vitro system, not self-replicate. This low-cost, easily propagated system provides a streamlined, instrument-free solution to improve the efficiency of TB diagnosis in resource-limited settings.

Lily Telisinghe of University Hospitals Plymouth NHS Trust together with Ben Swift at PBD Biotech Ltd in the United Kingdom will develop a system combining a biological agent and mechanical disruption for the rapid lysis of Mycobacterium tuberculosis from patient samples to enhance TB diagnosis. Tests will be performed to determine if there are constraints for two biosample types, tongue swabs and blood, on how soon after sampling they must be analyzed. These sample types will then be spiked with the BCG vaccine strain and used to determine the optimal combination of factors for cell lysis. This combination of conditions will then be used to test how the lysis system performs for TB diagnosis in patients in the United Kingdom and Indonesia.

Meng Sun of the Zymeron Corporation in the U.S. will develop a small, handheld, low-cost device for rapid plasma separation from whole blood for HIV diagnostic testing. Existing versions of the device will be modified to be able to process a larger volume of blood and deliver 100-200 microliters of plasma. The device is designed for untrained users. It can readily be modified to connect with blood drawing devices, including automatically sampling a fixed volume of blood to process, either by capillary action or direct loading, and delivering a fixed volume of plasma. It can also readily be integrated to deliver plasma to different diagnostic platforms, such as those based on microfluidics and lateral flow systems.

Salus Discovery in the U.S. will optimize a prototype of their simple and inexpensive SnapTab platform technology for processing of finger-prick blood for HIV diagnostic testing. SnapTab components and chemistry will be optimized for blood plasma separation, viral lysis, and in a final step, nucleic acid purification and stabilization. The output can then be directly used in standard quantitative PCR amplification reactions for HIV detection. To evaluate the performance of SnapTab for HIV, sample extraction/purification results will be compared against existing approaches including the plasma separation card and traditional bead-based processing of plasma.

Jeffrey Burke of Prompt Diagnostics, Inc. in the U.S. will develop a low-cost, automated platform integrating blood sample input, plasma separation, and HIV RNA extraction for HIV diagnostic testing. The platform will be based on magnetofluidic cartridge technology in which sequential bioassay steps are conducted via transfer of magnetic beads between reagents: red blood cell-binding beads in the plasma separation step and RNA-binding beads in the virion lysis step. A prototype cartridge will be built and optimized, including reagents in shelf-stable formats, and a low-cost, battery-powered device will be built to house the cartridge and drive the transfer of magnetic beads. The prototype platform will be tested for RNA quality and extraction efficiency, using blood samples spiked with HIV virions and comparing directly to standard clinical laboratory procedures.

Rainer Ng of Baebies, Inc. in the U.S. will develop a system for finger-prick blood self-collection and sample processing that yields a sample ready for HIV diagnostic testing either at the point of care or after delivery to a central laboratory. A simple, disposable sample collection device will incorporate a membrane for plasma separation. Squeezing the device delivers plasma to a tube in which HIV-binding magnetic beads concentrate virions and reagents stabilize them. The prototype system will be optimized to ensure it generates over 100 microliters of plasma from self-collected blood, captures virions of sufficient quality and quantity to enable standard RT-PCR testing, and stabilizes virions sufficiently for diagnostic testing over three days after sample collection.