Intra‐ and inter‐laboratory agreement of the FAM19A4/mir124‐2 methylation test: Results from an international study

BACKGROUND: HPV‐based cervical screening detects women at an increased risk of cervical cancer and precancer. To differentiate among HPV‐positive women those with (pre)cancer, triage testing is necessary. The detection of cancer‐associated host‐cell DNA methylation (FAM19A4 and hsa‐mir124‐2) in cervical samples has shown valuable as triage test. This multicenter study from 6 collaborating European laboratories and one reference laboratory was set out to determine the intra‐ and inter‐laboratory agreement of FAM19A4/mir124‐2 DNA methylation analysis utilizing the QIAsure Methylation Test. METHODS: Agreement analysis for the QIAsure Methylation Test was assessed on high‐risk HPV‐positive cervical specimens (n = 1680) both at the level of the assay and at the full workflow, including bisulfite conversion. RESULTS: Intra‐ and inter‐laboratory assay agreement were 91.4% (534/584; 95% CI 88.9‐93.5; κ = 0.82) and 92.5% (369/399; 95% CI 90.0‐94.7; κ = 0.83), respectively. The inter‐laboratory workflow (bisulfite conversion and assay combined) agreement was 90.0% (627/697; 95% CI 87.5%‐92.0%; κ = 0.76). CONCLUSION: These data show that the QIAsure Methylation Test performs robust and reproducible in different laboratory contexts. These results support the use of the QIAsure Methylation Test for full molecular screening for cervical cancer, including primary HPV testing and triage testing by methylation analysis.