Fig. 3

Results of the second round of single RPA primer screening for various HPV subtypes. a, b, d Amplification of plasmids of HPV types 16, 18, and 33 at different concentrations using the HF5-1-1/HR4 primer combination. c Amplification of plasmids of HPV type 31 at different concentrations using the HF5-1-3/HR4 primer combination. e Amplification of plasmids of HPV type 35 at different concentrations using the HF5-3-2/HR4 primer combination. f Amplification of plasmids of HPV type 45 at different concentrations using the HF5-2-2/HR4 primer combination. The forward primers in the above combinations share the same reverse primer, HR4. g Amplification of plasmids of HPV type 33 at different concentrations using the HF5-3-2/HR4-33Q2 primer combination. h Amplification of plasmids of HPV type 18 at different concentrations using the HF5-18Y/HR4-18Q2 primer combination. C1–C5: Plasmid concentrations of 10⁴ copies/μL, 10³ copies/μL, 10² copies/μL, 10 copies/μL, and 1 copy/μL respectively. N Negative control. The RPA amplification sensitivity for HPV types 18, 31, 33, and 35 is achieved at 1 copy/μL, while for HPV types 16 and 45, it is achieved at 10 copies/μL