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. 2021 Jun 17;11:12800. doi: 10.1038/s41598-021-92329-2

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Evaluation of the PGMY/GP6+ primer pool. (A) Schematic showing the relative position of the primer sets locating in the conservative L1 region of HPV. (B) Plasmids with different copy numbers of HPV16 and HPV18 were amplified by RPA reaction using the PGMY/GP6+ primer pool. NTC no-template control. (C) Real-time fluorescence detection of HPVs with the crRNA pool containing 11 crRNAs. For a single reaction, 5 μg of each individual HPV plasmid was used. NTC no-template control. (D) The PGMY/GP6+ pool was tested using 10,000 copies of the indicated HPV plasmids for each RPA reaction. The RPA reaction was performed at 37 °C for 20 min and followed by Cas12a detection at 37 °C for up to 60 min. NTC no-template control. (E) Nucleotide sequence alignments of GP6+ (positions 5879 to 5903 according to HPV16 sequence, Genbank accession number NC_001526.4) to 13 HR-HPV genotypes. HPV genotypes are specified by numbers on the left. Dots indicate the presence of nucleotides identical to the top sequence (GP6+). HPV Types with weak amplification by the primer pool are shown by box.