He Anna, Fang Cheng, Ming Yue, Tan He, Zhang Mengyi, Zhang Ruiqing, Li Jingyi, Nie Mingzhu, Li Fengyu, Hu Yaxin, Shen Xinxin, Rong Xiuge, Ma Xuejun
North China University of Science and Technology, Tangshan, 063210, China.
Tangshan Gongren Hospital, Tangshan, 063003, Hebei, China.
Heliyon. 2022 Nov 2;8(11):e11323. doi: 10.1016/j.heliyon.2022.e11323. eCollection 2022 Nov.
Human papillomavirus (HPV) 6 and 11 are the two most common low-risk HPV subtypes, accounting for more than 90% of condyloma acuminatum. A simple, accurate and rapid screening method to be applied in community-level hospitals is in high demand.
Endogenous internally controlled recombinase-assisted amplification (EIC-RAA) assays for HPV6 and 11 were performed in a single closed-tube at 39 °C within 30 min. The sensitivity and specificity of EIC-RAA were examined using recombinant plasmids and pre-tested HPV DNA. A total of 233 clinical samples were collected, and the DNA was extracted by traditional multi-step extraction, or sample releasing agent, before analysis by EIC-RAA. For comparison, HPV detection via Quantitative real-time PCR (qPCR) was also performed.
The sensitivity of EIC-RAA analysis was 10 copies/reaction for HPV6, 100 copies/reaction for HPV11, and 100 copies/reaction for the human β-globin gene. No cross-reaction was observed with other HPV subtypes. Clinical performance of the EIC-RAA assay achieved a 100% of concordance rate with the commercial HPV qPCR kit. Further, the EIC-RAA assay achieved a 100% of concordance rate when using multi-step extracted DNA and sample releasing agent-processed DNA.
The EIC-RAA assay for HPV6 and 11 detection possesses the advantages of accuracy, simplicity and rapidity, and demonstrates great potential to be used in community-level hospitals for field investigation.
人乳头瘤病毒（HPV）6型和11型是两种最常见的低风险HPV亚型，占尖锐湿疣病例的90%以上。目前迫切需要一种简单、准确、快速且适用于社区医院的筛查方法。
采用内源性内控重组酶辅助扩增（EIC-RAA）法在39℃下于单个封闭管中30分钟内对HPV6和11进行检测。使用重组质粒和预先检测的HPV DNA检测EIC-RAA的灵敏度和特异性。共收集233份临床样本，通过传统多步提取或样本释放剂提取DNA，然后进行EIC-RAA分析。作为对照，同时采用定量实时PCR（qPCR）法检测HPV。
EIC-RAA分析对HPV6的灵敏度为10拷贝/反应，对HPV11为100拷贝/反应，对人β-珠蛋白基因为100拷贝/反应。未观察到与其他HPV亚型的交叉反应。EIC-RAA检测的临床性能与商用HPV qPCR试剂盒的符合率达100%。此外，使用多步提取DNA和样本释放剂处理的DNA时，EIC-RAA检测的符合率也达100%。
用于检测HPV6和11的EIC-RAA检测法具有准确、简单、快速的优点，在社区医院现场检测中具有巨大的应用潜力。
