A Rapid CRISPR-based Field Test for the Non-invasive Detection of the Fungal Causative Agent of White-nose Syndrome Active
White-nose syndrome has killed more than five million bats since its discovery in 2006. USGS is developing a non-invasive and user-friendly CRISPR white-nose syndrome biosensor to allow for quick detection of the pathogen that causes the disease.
The Science Issue and Relevance: The accessibility to Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based genetic tools has given rise to applications beyond site-directed genome editing to the precise detection of DNA and RNA. These tools include diagnostic detection of human disease causative agents, such as SARS-CoV-2 and malaria. Despite the technology being rapid and cost-effective, CRISPR tools have not been applied to wildlife disease surveillance. This study presents the first development of a CRISPR-based diagnostic tool for the detection of a wildlife pathogen. White-nose syndrome (WNS) is an infectious disease caused by the fungus Pseudogymnoascus destructans (Pd) that is lethal to some North American bat species and has killed more than five million bats since its discovery in 2006. Here, a non-invasive and user-friendly CRISPR WNS biosensor was developed to enable rapid, point of use detection of the Pd pathogen to efficiently inform WNS surveillance and management actions.
Methodology for Addressing the Issue: Monitoring of Pd using traditional polymerase chain reaction (PCR)-based methods requires costly instruments which are often not suitable for the field, technical expertise, and the elimination of environmental inhibitors that interfere with molecular amplification. CRISPR-based detection tools have been demonstrated to be as sensitive and specific as PCR methods, but their chemistry has proven more robust to environmental inhibitors of molecular detection.
The CRISPR WNS biosensor assay employs CRISPR/Cas12a nuclease cleavage with recombinase polymerase amplification (RPA) to detect Pd DNA from bat dermal swabs and guano samples without the need for DNA extraction and purification. This tool provides a user-friendly and rapid (less than one hour) assay that is capable of detecting down to 158 copies of the target Pd DNA.
Future Steps: This CRISPR biosensor is a user-friendly and cost-effective tool that can be assembled and used by stakeholders with limited financial and logistical resources. Moreover, since the versatility of CRISPR-based diagnostics is owed to the simplicity of its targeting principles; new organisms of interest can be detected by simply exchanging the nucleic acid components that modulate target recognition.
White-nose syndrome has killed more than five million bats since its discovery in 2006. USGS is developing a non-invasive and user-friendly CRISPR white-nose syndrome biosensor to allow for quick detection of the pathogen that causes the disease.
The Science Issue and Relevance: The accessibility to Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based genetic tools has given rise to applications beyond site-directed genome editing to the precise detection of DNA and RNA. These tools include diagnostic detection of human disease causative agents, such as SARS-CoV-2 and malaria. Despite the technology being rapid and cost-effective, CRISPR tools have not been applied to wildlife disease surveillance. This study presents the first development of a CRISPR-based diagnostic tool for the detection of a wildlife pathogen. White-nose syndrome (WNS) is an infectious disease caused by the fungus Pseudogymnoascus destructans (Pd) that is lethal to some North American bat species and has killed more than five million bats since its discovery in 2006. Here, a non-invasive and user-friendly CRISPR WNS biosensor was developed to enable rapid, point of use detection of the Pd pathogen to efficiently inform WNS surveillance and management actions.
Methodology for Addressing the Issue: Monitoring of Pd using traditional polymerase chain reaction (PCR)-based methods requires costly instruments which are often not suitable for the field, technical expertise, and the elimination of environmental inhibitors that interfere with molecular amplification. CRISPR-based detection tools have been demonstrated to be as sensitive and specific as PCR methods, but their chemistry has proven more robust to environmental inhibitors of molecular detection.
The CRISPR WNS biosensor assay employs CRISPR/Cas12a nuclease cleavage with recombinase polymerase amplification (RPA) to detect Pd DNA from bat dermal swabs and guano samples without the need for DNA extraction and purification. This tool provides a user-friendly and rapid (less than one hour) assay that is capable of detecting down to 158 copies of the target Pd DNA.
Future Steps: This CRISPR biosensor is a user-friendly and cost-effective tool that can be assembled and used by stakeholders with limited financial and logistical resources. Moreover, since the versatility of CRISPR-based diagnostics is owed to the simplicity of its targeting principles; new organisms of interest can be detected by simply exchanging the nucleic acid components that modulate target recognition.