Analysis of the impact of non-matching nucleic acid extraction reagents on human papillomavirus genotyping results

Wu Litao; Zhang Wei; Zhou Hangyu; Zhang Mei; Shang E; Yan Hongchao; Liu Jiaxi; Guo Nan

doi:10.3760/j.issn.1673-4777.2026.01.017

Abstract：Objective To explore the impact of non-matching nucleic acid extraction reagents on human papillomavirus(HPV)genotyping results.Methods A total of 171 cervical exfoliated cell samples were collected from female patients who visited the Department of Gynecology,Dongzhimen Hospital,Beijing University of Chinese Medicine from March to April 2025.HPV DNA was extracted by manual boiling method and magnetic bead method(automatic nucleic acid extraction using non-matching nucleic acid extraction reagents),respectively.Fluorescence quantitative polymerase chain reaction was performed to detect HPV genotypes.The negative coincidence rate,positive coincidence rate,and total coincidence rate of nucleic acid detection results obtained by different extraction methods were analyzed,as well as their impacts on single infection,multiple infections of HPV subtypes,and sample viral load.Results The negative coincidence rate and positive coincidence rate of the detection results by the two methods were 100.0％(128/128)and 93.0％(40/43),respectively,with a total coincidence rate of 97.1％(166/171).Consistency analysis of gene subtypes and internal reference in 43 positive samples showed that the automatic nucleic acid extraction method did not cause a significant increase in the internal reference Ct value due to the reduced sample volume.Kendall's concordance coefficient test showed statistical significance(both P＜0.001),indicating consistency between the two extraction methods.The Kendall's concord-ance coefficients were 0.953 and 0.943,respectively,suggesting a strong degree of consistency.Analysis of the viral load of samples extracted by the two methods showed that consistent results and an increase of 1 order of magnitude accounted for 84.0％(42/50).Conclusion Compared with the manual boiling method,the magnetic bead method(automatic nucleic acid extraction with non-matching reagents)can obtain HPV DNA solutions with higher concentration and purity,which is more suitable for laboratory promotion and application.