Human papilloma virus (HPV) is the primary causative agent of cervical, vaginal, and vulvar cancers. HPV E6/E7 mRNA detection has been proven to improve the specificity and positive predictive value compared with HPV DNA testing in screening, whereby, it may possess higher diagnostic potential. Herein, to establish the ultrasensitive and specific detection of HPV E6/E7 mRNA, we developed a novel triple signal amplification strategy, combined with gold nanoparticles (AuNPs), reverse transcription loop-mediated isothermal amplification (RT-LAMP) and high affinity biotin-avidin system. This novel proposed signal amplification strategy exhibits the desired detection limit of 0.08 fM (approximately 100 copies) and a wide linear range from 0.1 pmol/mL to 100 nmol/mL for HPV16 E6/E7 mRNA detection. Importantly, the present novel biosensor is 10–100 times more sensitive than conventional RT-PCR in detecting HPV16 E6/E7 mRNA positive clinical samples. Conclusively, this biosensor shows good stability, selectivity, and reproducibility, which demonstrates its potential in future clinical diagnosis with desirable sensitivity and specificity.

人乳头瘤病毒 (HPV) 是宫颈癌、阴道癌和外阴癌的主要病原体。与HPV DNA检测相比，HPV E6/E7 mRNA检测已被证明可以提高筛查的特异性和阳性预测值，从而可能具有更高的诊断潜力。在此，为了建立 HPV E6/E7 mRNA 的超灵敏和特异性检测，我们开发了一种新的三重信号放大策略，结合金纳米粒子 (AuNPs)、逆转录环介导的等温扩增 (RT-LAMP) 和高亲和力生物素-亲和素系统。这种新提出的信号放大策略表现出 0.08 fM（大约 100 个拷贝）的所需检测限和 0.1 pmol/mL 至 100 nmol/mL 的宽线性范围，用于 HPV16 E6/E7 mRNA 检测。重要的，本新型生物传感器在检测 HPV16 E6/E7 mRNA 阳性临床样本方面比传统 RT-PCR 敏感 10-100 倍。总之，该生物传感器显示出良好的稳定性、选择性和重现性，这证明了其在未来临床诊断中的潜力，具有理想的灵敏度和特异性。