期刊
EXPERIMENTAL BIOLOGY AND MEDICINE
卷 246, 期 6, 页码 654-666出版社
SAGE PUBLICATIONS LTD
DOI: 10.1177/1535370220978899
关键词
Cell-free DNA; human papillomavirus; cervical cancer; droplet digital PCR; recombinase polymerase amplification; lateral flow strip
资金
- National Nanotechnology Center, Thailand [P1750034, P1851699, P1750162]
The study suggests that HPV E7 cfDNA could serve as a specific biomarker for cervical cancer detection with potential for monitoring. Both isothermal detection and ddPCR showed high sensitivity and specificity for HPV16/18 E7 cfDNA, while the RPA-LF method offers an affordable, rapid, and ultrasensitive tool for detecting HPV in resource-limited settings.
Circulating cell-free DNA (cfDNA) has attracted attention as a non-invasive biomarker for diagnosing and monitoring various cancers. Given that human papillomavirus (HPV) DNA integration and overexpression of E6/E7 oncogenes are pivotal events for carcinogenesis, we sought to determine if HPV E7 cfDNA could serve as a specific biomarker for cervical cancer detection. We applied droplet digital PCR (ddPCR) to quantify HPV16/18 E7 cfDNA from the serum of patients with cervical cancer, cervical intraepithelial neoplasia, and controls. HPV16/18 E7 cfDNA was highly specific for cervical cancer, displaying 30.77% sensitivity, 100% specificity, and an area under the curve of 0.65. Furthermore, we developed a sensitive isothermal detection of HPV16/18 E7 and the PIK3CA WT reference gene based on recombinase polymerase amplification combined with a lateral flow strip (RPA-LF). The assay took less than 30 min and the detection limit was 5-10 copies. RPA-LF exhibited 100% sensitivity and 88.24% specificity towards HPV16/18 E7 cfDNA in clinical samples. The agreement between RPA-LF and ddPCR was 83.33% (kappa = 0.67) for HPV16 E7 and 100% (kappa = 1.0) for HPV18 E7, indicating a good correlation between both tests. Therefore, we conclude that HPV E7 cfDNA represents a potential tumor marker with excellent specificity and moderate sensitivity for minimally invasive cervical cancer monitoring. Moreover, the RPA-LF assay provides an affordable, rapid, and ultrasensitive tool for detecting HPV cfDNA in resource-limited settings.
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