Internally controlled recombinase-aided amplification (IC-RAA) assays for the detection of human papillomavirus genotype
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ORIGINAL ARTICLE
Internally controlled recombinase‑aided amplification (IC‑RAA) assays for the detection of human papillomavirus genotypes 16 and 18 using extracted DNA and samples treated with nucleic acid releasing agent Jinrong Wang1,2,3 · Jianli Liu4 · Guowei Song3 · Zhi Cao5 · Jing Pan3 · Xinna Li2 · Yuan Gao1,2 · Juju Qi3 · Ziwei Chen2 · Guohao Fan2 · Xueding Bai2 · Ruiqing Zhang1,2 · Ruihuan Wang2 · Qingxia Duan1,2 · Lixin Li1,3 · Xinxin Shen2 · Xuejun Ma2 Received: 17 March 2020 / Accepted: 25 May 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract Cervical cancer is primarily caused by persistent infection with high-risk human papillomavirus (HPV), and 70% of cases are associated with HPV16 and 18 infections. The objective of this study was to establish rapid, simple, and sensitive internally controlled recombinase-aided amplification (IC-RAA) assays for the detection of HPV16 and 18. The assays were performed at 39 ℃ and were completed within 30 min. A total of 277 clinical samples of exfoliated cervical cells were tested by ICRAA assays and commercial HPV real-time fluorescent PCR kits using extracted DNA and samples treated with nucleic acid releasing agent. The analytical sensitivity of the IC-RAA assay was found to be 10 copies/μL for the detection of HPV16 and 18 when using recombinant plasmids as targets. The optimal concentration of the internal control (IC) plasmid and 18 was 1000 copies/μL for HPV16 and 100 copies/μL for HPV18. The clinical sensitivity of the IC-RAA assays for HPV16 using extracted DNA and samples treated with nucleic acid releasing agent was 98.73% and 97.47%, respectively, with kappa values of 0.977 (P
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