Research and Development Centre, Hangzhou D.A. Medical Laboratory, Hangzhou, Zhejiang, PR China.
J Med Microbiol. 2018 Oct;67(10):1509-1514. doi: 10.1099/jmm.0.000824. Epub 2018 Aug 20.
Human papillomavirus (HPV) E6/E7 mRNA is a more specific marker for cervical lesion screening than HPV DNA. Here, we aimed to develop a new one-step multiplex reverse transcript real-time PCR (MRT-PCR) to detect E6/E7 mRNA from 14 high-risk HPV (hrHPV) genotypes.
The analytical sensitivity and specificity of the MRT-PCR system were validated. Its clinical performance was evaluated by comparing the results with bDNA signal amplification assay and histopathological results.
The detection limit of MRT-PCR was 20 to 200 copies per reaction of different HPV genotypes, and no cross-reactivity was observed with any other low-risk HPV or other pathogens commonly found in the female genital tract. Using the bDNA signal amplification assay for comparison, a test on 166 clinical samples showed that the overall agreement between the two methods was 95.18 % and the one-step MRT-PCR was more sensitive. Further, compared with the histopathological results for the 166 clinical samples, the sensitivity and specificity of the MRT-PCR method were 88.9 and 71.6 %, respectively, and the positive rate for hrHPV E6/E7 mRNA increased with the severity of the cervical lesions.
The one-step multiplex RT-PCR for E6/E7 mRNA detection is a simple, fast, universally applicable, sensitive and highly specific method for hrHPV E6/E7 mRNA detection. It is reliable for cervical lesion screening and of potential value in future clinical applications.
人乳头瘤病毒（HPV）E6/E7 mRNA 是一种比 HPV DNA 更具特异性的宫颈病变筛查标志物。本研究旨在开发一种新的一步法多重逆转录实时 PCR（MRT-PCR），以检测来自 14 种高危型 HPV（hrHPV）的 E6/E7 mRNA。
验证了 MRT-PCR 系统的分析灵敏度和特异性。通过与 bDNA 信号扩增检测和组织病理学结果进行比较，评估其临床性能。
MRT-PCR 的检测限为 20 至 200 个拷贝/反应的不同 HPV 基因型，与任何其他低危型 HPV 或女性生殖道中常见的其他病原体均无交叉反应。使用 bDNA 信号扩增检测进行比较，对 166 例临床样本的检测表明，两种方法之间的总体一致性为 95.18%，一步法 MRT-PCR 更敏感。此外，与 166 例临床样本的组织病理学结果相比，MRT-PCR 方法的灵敏度和特异性分别为 88.9%和 71.6%，hrHPV E6/E7 mRNA 的阳性率随宫颈病变的严重程度而增加。
用于检测 E6/E7 mRNA 的一步法多重 RT-PCR 是一种简单、快速、通用、敏感且高度特异的检测 hrHPV E6/E7 mRNA 的方法。它可用于宫颈病变筛查，在未来的临床应用中具有潜在价值。
