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A research team led by Jennifer Doudna, winner of the 2020 Nobel Prize in Chemistry and a professor at the University of California, Berkeley, used CRISPR gene editing technology to propose a method to detect the new coronavirus in just 5 minutes.The test method does not require expensive laboratory equipment to operate and can be used in doctor’s offices, schools, and office buildings. Related results were published on the medRxiv preprint platform.
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Max Wilson, a molecular biologist at the University of California, Santa Barbara, said this seems like an absolutely reliable test.
In May of this year, two research teams reported on a new CRISPR-based coronavirus detection method, which can detect the virus in about 1 hour, much faster than the 24 hours required by traditional detection methods.
The recently proposed detection method is currently the fastest diagnostic method based on CRISPR.
The working principle of CRISPR detection is to recognize an RNA sequence of approximately 20 bases, which is a unique base sequence of the new coronavirus.
They created a “guide” RNA complementary to the target RNA sequence and combined with the target RNA sequence in solution.
When the “guide” RNA binds to the target RNA, the Cas13 “scissors” enzyme from the CRISPR tool will start and cut the nearby single-stranded RNA.
In addition, shearing will release individual fluorescent particles into the test solution.When the sample is irradiated with a laser, the released fluorescent particles will glow, indicating the presence of the virus.
The original CRISPR detection method requires researchers to first amplify the viral RNA and then perform detection and diagnosis, which undoubtedly increases the complexity, cost and time of detection. This new CRISPR diagnostic method does not require amplification of the new coronavirus RNA.
The team also spent several months testing hundreds of “guide” RNAs to find multiple “guide” RNAs that can work together to improve detection sensitivity.
The researchers report that using a single “guide” RNA, 100,000 viruses can be detected per microliter of solution. If they add a second “guide” RNA, they can detect 100 viruses per microliter.
Melanie Ott, a virologist at the University of California, San Francisco, who co-led the research, said the method is not yet as good as traditional novel coronavirus diagnostic devices, which use expensive laboratory machines to track viruses, which can track one virus per microliter.
However, he said the new diagnostic method can accurately identify a batch of 5 positive clinical samples, and each test takes just 5 minutes, while standard tests require 1 day or more to get results.
Wilson said the new detection method has another key advantage: It can quantify the amount of virus in a sample.
When traditional tests use the amplification of genetic material to achieve the purpose of detection, the amount of existing genetic material will change, making it impossible to determine the amount of virus in the sample.
In contrast, the intensity of the fluorescent signal detected by the new detection method is directly proportional to the number of viruses in the sample. This not only reveals whether the sample is positive, but also how much virus the patient is carrying.
Wilson said this information can help doctors make treatment plans based on each patient’s condition.
