A Novel Tailed Primer Nucleic Acid Test for Detection of HPV 16, 18 and 45 DNA at the Point of Care

Authors

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

Language

English

Publication Date

11-21-2023

Journal

Scientific Reports

DOI

10.1038/s41598-023-47582-y

PMID

37989845

PMCID

PMC10663460

PubMedCentral® Posted Date

11-21-2023

PubMedCentral® Full Text Version

Post-print

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.

Keywords

Human papillomavirus 16, Reproducibility of Results, DNA, Viral, Female, Point-of-Care Systems, Humans, Papillomavirus Infections, Genotype, Uterine Cervical Neoplasms, Papillomaviridae, Early Detection of Cancer, Human papillomavirus 18, Nucleic Acids, Cancer screening, Biomedical engineering

Published Open-Access

yes

Recommended Citation

Chang, Megan M; Ma, Ariel; Novak, Emilie Newsham; et al., "A Novel Tailed Primer Nucleic Acid Test for Detection of HPV 16, 18 and 45 DNA at the Point of Care" (2023). Center for Medical Ethics and Health Policy Staff Publications. 190.
https://digitalcommons.library.tmc.edu/med_ethics/190