Performance Analysis of Human Papillomavirus Viruses Genotype Detection Kit Using ZIP Nucleic Acid

초록

Objectives: This study aimed to evaluate the analytical and clinical performance of the MolecuTechⓇ Real HPV 16/18/HR kit, a ZIP nucleic acid (ZNA)-based c real-time polymerase chain reaction assay, for detecting human papillomavirus (HPV) genotypes linked to cervical cancer, and to confirm its reliability and enhanced annealing temperature (Tm) stability compared to the Anyplex II HPV28 Detection kit. Methods: Standard HPV DNA and 692 clinical samples from Korean hospitals were tested for limit of detection (LoD), reproducibility, specificity, sensitivity, and concordance with Anyplex II HPV28 Detection. The kit targets HPV 16, 18, and 12 highrisk genotypes (31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) using ZNA-enhanced primers and probes. Analytical validation used SiHa and HeLa cell lines, with reproducibility assessed over 60 replicates. Cohen’s kappa was calculated for concordance. Results: The LoD was 50 copies/reaction for HPV 16/18 and 100 copies/reaction for the 12 high-risk genotypes, with 40 cells/ mL (SiHa) and 8 cells/mL (HeLa) for cellular samples. Reproducibility showed a coefficient of variation <10% across 60 replicates, and specificity was 100% against 18 non-HPV microbes and 27 non-target HPV subtypes. Clinical sensitivity and specificity were 100% (95% confidence interval: 0.89–1.00 and 0.95–1.00, respectively), with 100% concordance (kappa >0.8) versus Anyplex II HPV28 Detection. Conclusion: The MolecuTechⓇ Real HPV 16/18/HR kit offers high sensitivity, specificity, and reproducibility, outperforming traditional assays in Tm stability due to ZNA technology, making it a reliable tool for HPV genotyping in clinical settings.

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