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CRISPR Test Diagnoses Mpox Faster Than Lab-Based PCR Method

By LabMedica International staff writers
Posted on 14 Feb 2024
Image: CRISPR is combined with nanopore sensing technology to detect whether or not mpox is present in a sample (Photo courtesy of 123RF)
Image: CRISPR is combined with nanopore sensing technology to detect whether or not mpox is present in a sample (Photo courtesy of 123RF)

Mpox, formerly known as monkeypox, is a rare viral disease transmitted through physical contact and typically presents mild symptoms like fever, rash, and swollen lymph nodes, though severe cases can require medical intervention. Due to its contagious nature, prompt testing is crucial for isolation and treatment purposes. Current mpox testing requires laboratory equipment and may take hours to yield results. Now, new research suggests a way for faster mpox testing that could be done in any clinic.

Researchers at Pennsylvania State University (University Park, PA, USA) have utilized CRISPR, the groundbreaking gene-editing technology, to develop a faster mpox test. For their study, the team designed a genetic sequence with an attached reporter specifically targeting the mpox virus. The test employs programmable CRISPR RNA that binds to the target and a protein called Cas12a, which together cleaves the reporter to produce fragments of varying sizes. Using nanopore sensing technology, these reporter fragments are analyzed to rapidly and accurately determine the presence of mpox in a sample.

The specificity of the test was validated by its inability to detect cowpox virus, a relative of mpox, thereby confirming its exclusive sensitivity to mpox. This testing method significantly reduces the detection time, taking only 32 to 55 minutes depending on the viral load, compared to the longer duration required for PCR lab testing. The research team is exploring the application of this nanopore technology for the development of tests for other pathogens, aiming to enable multi-target testing from a single sample using a portable device. Although this technology is not yet commercially available, the researchers hope to create a device that could facilitate widespread pathogen testing.

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Image: A research team led by Prof. Hsing I-Ming (center), Professor of the Department of Chemical and Biological Engineering at HKUST, has developed TEMPO, a thermodynamically programmed one-pot CRISPR platform for rapid, point-of-care nucleic acid testing. Mr. Wu Xiaolong (third right) and Ms. Li Yanan (first left), PhD students from the Department of Chemical and Biological Engineering, are the co-first authors of this study. (Photo courtesy of HKUST)

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