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New and improved probes may facilitate rapid verification of COVID-19 diagnostic tests

 


Nanoengineers at the University of California, San Diego have developed a new and improved probe called the Positive Control. This facilitates the validation of rapid point-of-care diagnostic tests for COVID-19 worldwide.

Positive controls made from virus-like particles are stable and easy to manufacture. Researchers say it has the potential to improve the accuracy of the new COVID-19 test, which is easier, faster, and cheaper to control, allowing the test to be extended outside the lab.

Our goal is to impact not only state-of-the-art hospitals, but also resource-poor and under-served areas that may lack advanced infrastructure and trained personnel. .. “

Nicole Steinmetz, Professor of Nanoengineering, University of California, San Diego, Jacobs Institute of Technology

Positive control is a staple of the lab. These are used to verify that the test or experiment actually works. The primary positive controls used to validate the COVID-19 test today are naked synthetic RNA, plasmids, or RNA samples from infected patients. But the problem is RNA, and the plasmid is not as stable as viral particles. They deteriorate easily, can require refrigeration, and are inconvenient and expensive to ship worldwide or store for extended periods of time.

In a paper published on November 25 ACS nanoResearchers at the University of California, San Diego, led by Steinmetz, report that packaging RNA segments from the SARS-CoV-2 virus into virus-like particles can create a stable positive control for the COVID-19 test. I am. It retains 70% of its activity after storage for one week and one month at temperatures up to 40 C (104 F) and can pass detection as a novel coronavirus without infection.

The team has developed two different controls. One was made from plant virus nanoparticles and the other was made from bacteriophage nanoparticles. Their use is easy. The controls are run and analyzed with a sample of the patient to provide a reliable benchmark of what a positive test result will look like.

To create plant virus-based controls, researchers use cowpea chlorotic mottle virus, which infects cowpea plants. They essentially open the virus, remove its RNA content, replace them with a synthesized RNA template containing specific sequences from the SARS-CoV-2 virus, and close everything.

The process of creating a bacteriophage-based control begins with a plasmid, which is a ring of DNA. These plasmids contain the gene sequence of interest for the SARS-CoV-2 virus and the gene encoding the surface protein of bacteriophage Qbeta. These plasmids are incorporated into the bacterium. This process reprograms the bacteria to produce virus-like particles with the SARS-CoV-2 RNA sequence on the inside and the Qbeta bacteriophage protein on the outside.

Both controls were validated in clinical samples. The big advantage researchers point out is that they can be used at every step of the COVID-19 test, unlike the positive controls used today.

“These can be used as complete process controls. The analysis can be performed in parallel with patient samples starting with RNA extraction,” said Soo Khim Chan, a postdoctoral researcher in Steinmetz’s lab. .. “Usually other controls will be added in a later step, so if you have a problem in the first step, you won’t know it.”

So far, researchers have adapted the controls for use in CDC-certified RT-PCR tests. This is currently the gold standard for COVID-19 testing, but it can take days to return results due to the costly, complex, and logistics of sending samples to laboratories with PCR capabilities.

Steinmetz, Chan and colleagues are currently working on adapting controls for use in less complex diagnostic tests, such as the RT-LAMP test, which can be run out-of-lab and provide immediate results.

“Making low-tech assays more accurate is a relatively simple nanotechnology approach,” Steinmetz said. “This can help break down some barriers to mass testing of underserved populations in the United States and around the world.”

Source:

Journal reference:

Chan, SK, et al. (2020) Severe Acute Respiratory Syndrome Coronavirus 2 Biomimetic virus-like particles as a diagnostic tool. ACS Nano. doi.org/10.1021/acsnano.0c08430..

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