Using gene editing proteins combined with nanopore sensing technology, new research shows that testing for mpox pathogens could become faster and more accessible.
Mpox (previously known as monkeypox) is a rare viral disease that is spread through physical contact between people. Currently, testing for mpox requires laboratory equipment and can take a few hours to get test results.
Although mpox symptoms are generally mild with fever, rash and swollen lymph nodes, severe cases can occur and require medical attention. Because the disease is contagious, testing is important so that people with the disease can isolate until symptoms resolve or get appropriate medical care.
A new study, carried out by Ahasan Ahamed MD, a graduate student at Pennsylvania State University, used CRISPR technology to create a faster diagnostic test. Since 2017, scientists have expanded the application of CRISPR technology from gene editing to molecular diagnostic techniques.
For this study, Ahamed created a genetic sequence combined with a reporter to specifically target the mpox virus. Then a programmable CRISPR RNA binds to both the target and a protein called Cas12a and, together, the CRISPR/Cas12a cleaves the reporter to create various sizes of fragments. The researchers can then use nanopore sensing technology to analyse those reporters’ fragments, providing a rapid and accurate test that detects whether or not mpox is present in the sample.
The team confirmed that the test they created is specific to mpox; when they tested samples of cowpox virus, a close relative of mpox, the test did not show a positive result.
The whole process is quick, taking 32 to 55 minutes to detect the target, depending on viral load. Much faster than it currently takes to test for mpox in a laboratory using PCR method. The research was presented at the recent 68th Biophysical Society Annual Meeting.
The researchers plan to apply this nanopore technology to create tests for other pathogens, allowing one sample to be tested for multiple targets using portable device. And while the technology is not currently commercially available, the research authors are hopeful that they will soon create a device that could make this kind of pathogen testing widely available.