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CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics.
- Source :
-
ACS synthetic biology [ACS Synth Biol] 2024 Mar 15; Vol. 13 (3), pp. 837-850. Date of Electronic Publication: 2024 Feb 13. - Publication Year :
- 2024
-
Abstract
- The World Health Organization's global initiative toward eliminating high-risk Human Papillomavirus (hrHPV)-related cancers recommends DNA testing over visual inspection in all settings for primary cancer screening and HPV eradication by 2100. However, multiple hrHPV types cause different types of cancers, and there is a pressing need for an easy-to-use, multiplex point-of-care diagnostic platform for detecting different hrHPV types. Recently, CRISPR-Cas systems have been repurposed for point-of-care detection. Here, we established a CRISPR -Cas multiplexed d iagnostic assay (CRISPRD) to detect cervical cancer-causing hrHPVs in one reaction (one-pot assay). We harnessed the compatibility of thermostable AapCas12b, TccCas13a, and HheCas13a nucleases with isothermal amplification and successfully detected HPV16 and HPV18, along with an internal control in a single-pot assay with a limit of detection of 10 copies and 100% specificity. This platform offers a rapid and practical solution for the multiplex detection of hrHPVs, which may facilitate large-scale hrHPV point-of-care screening. Furthermore, the CRISPRD platform programmability enables it to be adapted for the multiplex detection of any two nucleic acid biomarkers as well as internal control.
- Subjects :
- Female
Humans
Human Papillomavirus Viruses
Point-of-Care Testing
Human papillomavirus 16 genetics
Papillomavirus Infections diagnosis
Papillomavirus Infections genetics
Papillomavirus Infections prevention & control
Uterine Cervical Neoplasms diagnosis
Uterine Cervical Neoplasms prevention & control
Subjects
Details
- Language :
- English
- ISSN :
- 2161-5063
- Volume :
- 13
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- ACS synthetic biology
- Publication Type :
- Academic Journal
- Accession number :
- 38349963
- Full Text :
- https://doi.org/10.1021/acssynbio.3c00655