CRISPR-based diagnostic technology for infectious diseases

A group of researchers from VISTEC, Siriraj hospital, Mahidol University and Department of Medical Sciences, Ministry of Public Health successfully developed a Thai FDA-approved test kit with CRISPR-based technology that can simultaneously detects 2 target genes in SARS-CoV-2 with high sensitivity and specificity. 

CRISPR-based diagnostics was designed for ease of use by integrating two major steps. The first step is to generate more copies of target nucleotide sequences using an isothermal amplification reaction, which we have the capacity to produce in-house. The second step is to detect amplified nucleic acids using corresponding Cas enzymes programmed with CRISPR RNAs. The resulting fluorescence signal can be detected either by naked eyes with blue LED pad illuminator or real-time PCR machine. The two-step reactions can be combined into a single tube for reduction of contamination and turnaround time while maintaining sensitivity and specificity for diagnostic purposes.

Figure 1 :  Dual-Gene CRISPR SARS-CoV-2 RNA detection kit

Beyond the scope in detection of human pathogens, CRISPR diagnostics can be developed for the detection of antimicrobial resistance, genetic disorders, agricultural pathogens, and circulating DNAs and RNAs associated with diseases. Another advantage of this technology is that multiple orthogonal Cas enzymes can work in symphony which would enable detection of multiple targets in only 1-2 tubes. To sum up, the CRISPR-based diagnostics has multiple advantages and rooms for further development for wide range of applications. The ultimate goal of the team is to ensure better accessibility to high-quality diagnostic tools for the Thai population, and to create broadly useful technologies that can be adapted worldwide.

Figure 2 : Summary of process from samples to results in Dual-Gene CRISPR SARS-CoV-2 RNA detection kit

Affiliation/Departments/Agencies:

• School of Biomolecular Science and Engineering, VISTEC
• Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University
• Department of Biochemistry, Faculty of Science, Mahidol University
• Department of Medical Sciences, Ministry of Public Health

Contact Us

Dr. Chayasith Uttamapinant (Team Leader)

School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC)

Wangchan Valley 555 Moo 1 Payupnai, Wangchan, 

Rayong 21210 Thailand

Email :  chayasith.u@vistec.ac.th