The nanopores are so tiny that they can partition the mixture into single droplets containing only one RNA molecule, which has the Cas13 protein attached. Once the RNA is combined with Cas13, the researchers place a nanopore polycarbonate membrane - a thin filter that is readily available, making it a cost-effective material, Guan said - on the mixture. They are using CRISPR-Cas13 to detect and signal the presence of HIV. In this study, the researchers are leveraging the technology not for its editing capabilities, but for its diagnostic potential. “We then mix this RNA with a special protein called Cas13, which is part of the CRISPR system.”ĬRISPR-Cas13 is a “revolutionary tool,” Guan said, that enables researchers to target and manipulate RNA sequences. “Here’s how it works: We take a small sample of a person’s blood and extract the viral RNA - the genetic material of the virus - from it,” Guan said. Guan and his team took a more direct approach with their test, named Self-digitization Through Automated Membrane-based Partitioning (STAMP), which is less expensive, quicker and requires less blood than the RT-PCR. The gold standard test, called RT-PCR, can produce a close estimate of the actual viral load, but it is not a direct measurement. They published the work in ACS Nano.Īccording to the corresponding author Weihua Guan, associate professor of electrical engineering and biomedical engineering in the Penn State College of Engineering, the digital assay is the first step in providing a clinical diagnostic tool for a bevy of infectious diseases.Ĭonventional tests for HIV viral load involve taking genetic material from the sample, amplifying it and comparing it to reference samples. Now, a Penn State research team has developed a time and cost-efficient digital assay that can directly measure the presence of HIV in a single drop of blood. The time-consuming viral load testing needs to be repeated several times as a patient undergoes treatment. Called the viral load, this is what is measured to allow clinicians to understand how patients are responding to anti-viral medications and monitor potential progression. For a person with human immunodeficiency virus (HIV), each drop of blood could contain anywhere from fewer than 20 copies of the virus to more than 500,000 copies. A milliliter of blood contains about 15 individual drops.
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