TY - JOUR
T1 - CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics
AU - Ghouneimy, Ahmed
AU - Ali, Zahir
AU - Aman, Rashid
AU - Jiang, Wenjun
AU - Aouida, Mustapha
AU - Mahfouz, Magdy
N1 - Publisher Copyright:
© 2024 The Authors. Published by American Chemical Society.
PY - 2024/3/15
Y1 - 2024/3/15
N2 - 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 diagnostic 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.
AB - 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 diagnostic 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.
KW - CRISPR-Cas
KW - cervical cancer
KW - human papillomavirus
KW - molecular diagnostics
KW - multiplex detection
KW - point-of-care detection
KW - sexually transmitted infections
UR - http://www.scopus.com/inward/record.url?scp=85185608945&partnerID=8YFLogxK
U2 - 10.1021/acssynbio.3c00655
DO - 10.1021/acssynbio.3c00655
M3 - Article
C2 - 38349963
AN - SCOPUS:85185608945
SN - 2161-5063
VL - 13
SP - 837
EP - 850
JO - ACS Synthetic Biology
JF - ACS Synthetic Biology
IS - 3
ER -