A novel tailed primer nucleic acid test for detection of HPV 16, 18 and 45 DNA at the point of care
Abstract Cervical cancer is a leading cause of death for women in low-resource settings despite being preventable through human papillomavirus (HPV) vaccination, early detection, and treatment of precancerous lesions. The World Health Organization recommends high-risk HPV (hrHPV) as the preferred ce...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-11-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-47582-y |
| _version_ | 1797453152685916160 |
|---|---|
| author | Megan M. Chang Ariel Ma Emilie Newsham Novak Maria Barra Kathryn A. Kundrod Jane Richards Montealegre Michael E. Scheurer Philip E. Castle Kathleen Schmeler Rebecca Richards-Kortum |
| author_facet | Megan M. Chang Ariel Ma Emilie Newsham Novak Maria Barra Kathryn A. Kundrod Jane Richards Montealegre Michael E. Scheurer Philip E. Castle Kathleen Schmeler Rebecca Richards-Kortum |
| author_sort | Megan M. Chang |
| collection | DOAJ |
| description | Abstract Cervical cancer is a leading cause of death for women in low-resource settings despite being preventable through human papillomavirus (HPV) vaccination, early detection, and treatment of precancerous lesions. The World Health Organization recommends high-risk HPV (hrHPV) as the preferred cervical cancer screening strategy, which is difficult to implement in low-resource settings due to high costs, reliance on centralized laboratory infrastructure, and long sample-to-answer times. To help meet the need for rapid, low-cost, and decentralized cervical cancer screening, we developed tailed primer isothermal amplification and lateral flow detection assays for HPV16, HPV18, and HPV45 DNA. We translated these assays into a self-contained cartridge to achieve multiplexed detection of three hrHPV genotypes in a disposable cartridge. The developed test achieves clinically relevant limits of detection of 50–500 copies per reaction with extracted genomic DNA from HPV-positive cells. Finally, we performed sample-to-answer testing with direct lysates of HPV-negative and HPV-positive cell lines and demonstrated consistent detection of HPV16, HPV18, and HPV45 with 5000–50,000 cells/mL in < 35 min. With additional optimization to improve cartridge reliability, incorporation of additional hrHPV types, and validation with clinical samples, the assay could serve as a point-of-care HPV DNA test that improves access to cervical cancer screening in low-resource settings. |
| first_indexed | 2024-03-09T15:18:43Z |
| format | Article |
| id | doaj.art-99092a73ebd440dfb40715212f927462 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-09T15:18:43Z |
| publishDate | 2023-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-99092a73ebd440dfb40715212f9274622023-11-26T12:56:00ZengNature PortfolioScientific Reports2045-23222023-11-0113111510.1038/s41598-023-47582-yA novel tailed primer nucleic acid test for detection of HPV 16, 18 and 45 DNA at the point of careMegan M. Chang0Ariel Ma1Emilie Newsham Novak2Maria Barra3Kathryn A. Kundrod4Jane Richards Montealegre5Michael E. Scheurer6Philip E. Castle7Kathleen Schmeler8Rebecca Richards-Kortum9Department of Bioengineering, Rice UniversityDepartment of Bioengineering, Rice UniversityDepartment of Bioengineering, Rice UniversityDepartment of Bioengineering, Rice UniversityDepartment of Bioengineering, Rice UniversityDepartment of Behavioral Science, The University of Texas MD Anderson Cancer CenterDepartment of Pediatrics Hematology/Oncology, Baylor College of Medicine, Texas Children’s HospitalDivision of Cancer Epidemiology and Genetics, National Cancer InstituteDepartment of Gynecologic Oncology and Reproductive Medicine, The University of Texas MD Anderson Cancer CenterDepartment of Bioengineering, Rice UniversityAbstract Cervical cancer is a leading cause of death for women in low-resource settings despite being preventable through human papillomavirus (HPV) vaccination, early detection, and treatment of precancerous lesions. The World Health Organization recommends high-risk HPV (hrHPV) as the preferred cervical cancer screening strategy, which is difficult to implement in low-resource settings due to high costs, reliance on centralized laboratory infrastructure, and long sample-to-answer times. To help meet the need for rapid, low-cost, and decentralized cervical cancer screening, we developed tailed primer isothermal amplification and lateral flow detection assays for HPV16, HPV18, and HPV45 DNA. We translated these assays into a self-contained cartridge to achieve multiplexed detection of three hrHPV genotypes in a disposable cartridge. The developed test achieves clinically relevant limits of detection of 50–500 copies per reaction with extracted genomic DNA from HPV-positive cells. Finally, we performed sample-to-answer testing with direct lysates of HPV-negative and HPV-positive cell lines and demonstrated consistent detection of HPV16, HPV18, and HPV45 with 5000–50,000 cells/mL in < 35 min. With additional optimization to improve cartridge reliability, incorporation of additional hrHPV types, and validation with clinical samples, the assay could serve as a point-of-care HPV DNA test that improves access to cervical cancer screening in low-resource settings.https://doi.org/10.1038/s41598-023-47582-y |
| spellingShingle | Megan M. Chang Ariel Ma Emilie Newsham Novak Maria Barra Kathryn A. Kundrod Jane Richards Montealegre Michael E. Scheurer Philip E. Castle Kathleen Schmeler Rebecca Richards-Kortum A novel tailed primer nucleic acid test for detection of HPV 16, 18 and 45 DNA at the point of care Scientific Reports |
| title | A novel tailed primer nucleic acid test for detection of HPV 16, 18 and 45 DNA at the point of care |
| title_full | A novel tailed primer nucleic acid test for detection of HPV 16, 18 and 45 DNA at the point of care |
| title_fullStr | A novel tailed primer nucleic acid test for detection of HPV 16, 18 and 45 DNA at the point of care |
| title_full_unstemmed | A novel tailed primer nucleic acid test for detection of HPV 16, 18 and 45 DNA at the point of care |
| title_short | A novel tailed primer nucleic acid test for detection of HPV 16, 18 and 45 DNA at the point of care |
| title_sort | novel tailed primer nucleic acid test for detection of hpv 16 18 and 45 dna at the point of care |
| url | https://doi.org/10.1038/s41598-023-47582-y |
| work_keys_str_mv | AT meganmchang anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT arielma anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT emilienewshamnovak anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT mariabarra anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT kathrynakundrod anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT janerichardsmontealegre anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT michaelescheurer anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT philipecastle anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT kathleenschmeler anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT rebeccarichardskortum anoveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT meganmchang noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT arielma noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT emilienewshamnovak noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT mariabarra noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT kathrynakundrod noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT janerichardsmontealegre noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT michaelescheurer noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT philipecastle noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT kathleenschmeler noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare AT rebeccarichardskortum noveltailedprimernucleicacidtestfordetectionofhpv1618and45dnaatthepointofcare |