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New 3D-Printed Tech Lowers Cost of Common Medical Test.

A desire for a simpler, cheaper way to do common laboratory tests for medical diagnoses and to avoid "washing the dishes" led UConn researchers to develop a new technology that reduces cost and time. Their pipette-based technology could also help make certain medical testing available in rural or remote areas where traditional methods might otherwise be prohibitively expensive and complicated to conduct.

The 3D-printed pipette-tip test developed by the researchers leverages what "has long been the gold standard for measuring proteins, pathogens, antibodies and other biomolecules in complex matrices," they claim. The method still employs the enzyme-linked immunosorbent assay, also known as ELISA, but through a different route. They detailed their findings in a paper recently published online in Analytical Chemistry.

For 30 years or more, ELISA has been used to test blood, cells and other biological samples for everything from certain cancers to HIV, from Lyme disease to pernicious anemia. Traditional ELISA tests are performed on plates featuring 96 microwells; each well works as a separate testing chamber where samples can be combined with various agents that will react with the sample, typically by changing color. Technicians then analyze whether a sample contains indicators of a particular disease or condition depending on the intensity of the color produced during the reaction.

While effective and accurate, equipment used to run ELISA is expensive--often costing thousands of dollars to install--and requires specialized training to conduct testing, as improper techniques can lead to incorrect results. The agents used in the actual tests--usually various forms of antibodies--can also be expensive.

Like many research laboratories, James Rusling's chemistry lab where research assistant Mohamed Sharafeldin and his primary collaborator, Karteek Kadimisetty '18 Ph.D., conducted their work, doesn't have an automated ELISA washing machine, meaning plates being used for tests must be manually washed--a time consuming and difficult process.

"The ELISA washing techniques take forever," said Sharafeldin, who is currently working toward his doctorate in chemistry. "It's very tough, especially in a lab like ours. We don't have those kind of fancy washing machines."

When Kadimisetty was running ELISA one day, he mentioned, "I wish doing ELISA was as simple as pipetting." That offhand comment was the impetus for what followed: a design for a 3D-printed adapter for commonly used pipettes that could run an ELISA test right in the pipette tip, without the need for a traditional ELISA plate and the expensive equipment that goes with it.

Each single-use pipette tip represents one micro-well on an ELISA plate; the researchers also designed a multi-tipped version that allows for eight tips to be pipetted at the same time. The tips fit snugly onto most pipettes used in laboratory settings, making fluid handling much easier than with the standard ELISA plate.

"We didn't want to make a big change in the traditional ELISA; we just made engineered, controlled changes," Sharafeldin said. "So, the basics are the same. We use the same antibodies at the same concentrations they use with conventional or traditional ELISA, so we are using the same protocols. Anything that can be run by normal ELISA can be run by this, with the advantage of being less expensive, much faster and accessible."

The researchers tested the pipette tips on samples from prostate cancer patients and found the test results from the tips were as accurate as ELISA tests and could conduct the tests with one-tenth the amount of testing agent--significantly reducing overall cost--at a fraction of the time. Tests conducted by different users with different levels of skill demonstrated the same results.

Traditional ELISA plate micro-wells hold 400 microliters of fluids each, but the reactions needed to measure test results only occur on the plastic walls of the well. While the 3D-printed ELISA tips hold only 50 microliters, the design of the reservoir inside the tip dramatically increases the surface area where reactions occur, allowing the researchers to use much less of the costly antibodies used to conduct the test, and reducing the time needed to process the test and read the results.

"Here we have a chamber where the reaction happens at all points," Sharafeldin said, referring to the pipette tip design. "This reduces the time of the assay, which is an important thing, because the ELISA assay takes from five to eight hours to run. This one can be run in 90 minutes."

The pipette tips also don't require an expensive or sophisticated plate reader to determine test results, as ELISA tests do. In the trials with the prostate cancer samples, the pipette tip results were accurately read by taking a cell phone photo and using a free app that measures color intensities in the image.

The user conducting the test with the pipette tips doesn't have to be a scientist; they just need simple pipetting instructions, then to take a photograph and send it to a technician who could remotely read the results to help make a diagnosis--providing new, lower-cost testing options in rural or isolated areas where establishing a traditional ELISA lab is challenging and expensive.

While additional sample testing is needed, Sharafeldin is optimistic about the future potential for the pipette tip design to reduce costs. He is also engaging with engineers to design an automated, vacuum-assisted pipette that would further ease use of the pipette tips and conducting ELISA tests, and would be available for significantly less cost than traditional ELISA equipment.

DoD Awards Contracts to Create Autonomous Robotic Trauma Care System

The University of Pittsburgh School of Medicine and Carnegie Mellon University have been awarded four-year contracts totaling over $7.2 million from the U.S. Department of Defense to create an autonomous trauma care system that fits in a backpack and can treat and stabilize soldiers injured in remote locations.

The goal of "TRAuma Care In a Rucksack: TRACIR" is to develop AI technologies enabling medical interventions that extend the "golden hour" for treating combat casualties and ensure an injured person's survival for long medical evacuations.

A multidisciplinary team of Pitt researchers and clinicians from emergency medicine, surgery, critical care, and pulmonary fields will provide a wealth of real-world trauma data and medical algorithms. CMU roboticists and computer scientists will incorporate in the creation of a "hard and soft robotic suit" into which an injured person can be placed. Monitors embedded in the suit will assess the injury, and AI algorithms will guide the appropriate critical care interventions and robotically apply stabilizing treatments, such as intravenous fluids and medications.

Ron Poropatich, M.D., retired U.S. Army colonel, director of Pitt's Center for Military Medicine Research, and professor in Pitt's Division of Pulmonary, Allergy and Critical Care Medicine, is overall principal investigator on the $3.71 million Pitt contract, with Michael R. Pinsky, M.D., professor in Pitt's Department of Critical Care Medicine, as its scientific principal investigator. Artur Dubrawski, Ph.D., research professor at CMU's Robotics Institute, is principal investigator on the $3.5 million CMU contract.

"Battlefields are becoming increasingly remote, making medical evacuations more difficult," said Poropatich. "By fusing data captured from multiple sensors and applying machine learning, we are developing more predictive cardio-pulmonary resuscitation opportunities, which hopefully will conserve an injured soldier's strength. Our goal with TRACIR is to treat and stabilize soldiers in the battlefield, even during periods of prolonged field care, when evacuation is not possible."

Much technology still must be developed to enable robots to reliably and safely perform tasks like inserting IV needles or placing a chest tube in the field, Dubrawski said. Initially, the research will be "a series of baby steps," demonstrating the practicality of individual components the system will eventually require.

"Everybody has a slightly different vision of what the final system will look like," Dubrawski added. "But we see this as being an autonomous or nearly autonomous system--a backpack containing an inflatable vest or perhaps a collapsed stretcher you might toss toward a wounded soldier. It would then open up, inflate, position itself and begin stabilizing the patient. Whatever human assistance it might need could be provided by someone without medical training."

With a digital library of detailed physiologic data collected from over 5,000 UPMC trauma patients, Pinsky and Dubrawski previously created algorithms that could allow a computer program to "learn" the signals that an injured patient's health is deteriorating before damage is irreversible and tell the robotic system to administer the best treatments and therapies.

"Pittsburgh has the three components you need for a project like this--expertise in critical care medicine, artificial intelligence and robotics," Dubrawski said. "That's why Pittsburgh is unique and is the one place for this project."

While the immediate goal of the project is to carry forward the U.S. military's principle of "leave no man behind," and treat soldiers on the battlefield, there are numerous potential civilian applications, said Poropatich.

"TRACIR could be deployed by drone to hikers or mountain climbers injured in the wilderness; it could be used by people in submarines or boats; it could give trauma care capabilities to rural health clinics or be used by aid workers responding to natural disasters," he said. "And, someday, it could even be used by astronauts on Mars."

Caption: This unique pipette tip was created with a 3D printer. Image courtesy of Sean Flynn/UConn.
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Title Annotation:IN DEVELOPMENT
Publication:Medical Product Outsourcing
Date:Jun 1, 2019
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