CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics
<p dir="ltr">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 typ...
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2024
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| _version_ | 1864513510314082304 |
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| author | Ahmed Ghouneimy (17959417) |
| author2 | Zahir Ali (777469) Rashid Aman (775203) Wenjun Jiang (141985) Mustapha Aouida (417652) Magdy Mahfouz (777477) |
| author2_role | author author author author author |
| author_facet | Ahmed Ghouneimy (17959417) Zahir Ali (777469) Rashid Aman (775203) Wenjun Jiang (141985) Mustapha Aouida (417652) Magdy Mahfouz (777477) |
| author_role | author |
| dc.creator.none.fl_str_mv | Ahmed Ghouneimy (17959417) Zahir Ali (777469) Rashid Aman (775203) Wenjun Jiang (141985) Mustapha Aouida (417652) Magdy Mahfouz (777477) |
| dc.date.none.fl_str_mv | 2024-02-13T09:00:00Z |
| dc.identifier.none.fl_str_mv | 10.1021/acssynbio.3c00655 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/CRISPR-Based_Multiplex_Detection_of_Human_Papillomaviruses_for_One-Pot_Point-of-Care_Diagnostics/26355106 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biomedical and clinical sciences Clinical sciences Medical biotechnology Oncology and carcinogenesis CRISPR-Cas Human Papillomavirus Sexually Transmitted Infections Molecular Diagnostics Multiplex Detection Point-Of-Care Detection Cervical Cancer |
| dc.title.none.fl_str_mv | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <p dir="ltr">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.</p><h2>Other Information</h2><p dir="ltr">Published in: ACS Synthetic Biology<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.1021/acssynbio.3c00655" target="_blank">https://dx.doi.org/10.1021/acssynbio.3c00655</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_59b2e1d6636335ccbc0ea0f47821bd03 |
| identifier_str_mv | 10.1021/acssynbio.3c00655 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/26355106 |
| publishDate | 2024 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care DiagnosticsAhmed Ghouneimy (17959417)Zahir Ali (777469)Rashid Aman (775203)Wenjun Jiang (141985)Mustapha Aouida (417652)Magdy Mahfouz (777477)Biomedical and clinical sciencesClinical sciencesMedical biotechnologyOncology and carcinogenesisCRISPR-CasHuman PapillomavirusSexually Transmitted InfectionsMolecular DiagnosticsMultiplex DetectionPoint-Of-Care DetectionCervical Cancer<p dir="ltr">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.</p><h2>Other Information</h2><p dir="ltr">Published in: ACS Synthetic Biology<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.1021/acssynbio.3c00655" target="_blank">https://dx.doi.org/10.1021/acssynbio.3c00655</a></p>2024-02-13T09:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.1021/acssynbio.3c00655https://figshare.com/articles/journal_contribution/CRISPR-Based_Multiplex_Detection_of_Human_Papillomaviruses_for_One-Pot_Point-of-Care_Diagnostics/26355106CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/263551062024-02-13T09:00:00Z |
| spellingShingle | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics Ahmed Ghouneimy (17959417) Biomedical and clinical sciences Clinical sciences Medical biotechnology Oncology and carcinogenesis CRISPR-Cas Human Papillomavirus Sexually Transmitted Infections Molecular Diagnostics Multiplex Detection Point-Of-Care Detection Cervical Cancer |
| status_str | publishedVersion |
| title | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics |
| title_full | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics |
| title_fullStr | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics |
| title_full_unstemmed | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics |
| title_short | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics |
| title_sort | CRISPR-Based Multiplex Detection of Human Papillomaviruses for One-Pot Point-of-Care Diagnostics |
| topic | Biomedical and clinical sciences Clinical sciences Medical biotechnology Oncology and carcinogenesis CRISPR-Cas Human Papillomavirus Sexually Transmitted Infections Molecular Diagnostics Multiplex Detection Point-Of-Care Detection Cervical Cancer |