期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 379, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2022.133231
关键词
CRISPR; Cas12a; Methylated SEPT9; MSRE; RPA; Cervical cancer detection
Cervical cancer is a common tumor in women, and early diagnosis is crucial. However, there is a lack of specific biomarkers and efficient methods. In this study, methylated SEPT9 (mSEPT9) was identified as a specific biomarker, and a novel method called MeCRISPR was developed for rapid and ultrasensitive detection of mSEPT9. The results showed high sensitivity and specificity in detecting mSEPT9 in clinical cervical cancer patients. This method has the potential for cancer screening in low-resource settings.
Cervical cancer is the fourth most common tumor in women with high mortalities. Early diagnosis can improve the survival rate. Nevertheless, there is still a lack of specific biomarkers and efficient methods for the diagnosis of cervical cancer with excellent sensitivity and specificity. In this work, we identified the methylated SEPT9 (mSEPT9) as a specific biomarker for the diagnosis of cervical cancer. We developed a novel method using methylation-sensitive restriction endonuclease (MSRE) combined with recombinase polymerase amplification (RPA)-based CRISPR/Cas12a system (MeCRISPR) for rapid and ultrasensitive detection of mSEPT9. We demonstrated the MeCRISPR enabled to detect 1 copy/mu L of mSEPT9 and distinguishes 0.01% mSEPT9 from large amounts of gDNA. This method could be completed in 60 min without any complicated instruments, which is favorable for point-of-care testing (POCT). We applied MeCRISPR to detect mSEPT9 in clinical cervical cancer patients. The sensitivity and specificity were 100% and 92.3% respectively. Our results demonstrate that mSEPT9 is a promising biomarker for cervical cancer. The MeCRISPR is a sensitive and simple method to analyze the methylated gene which holds great potential for screening cancer in a low-resource setting.
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