Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literatureReport as inadecuate




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Aim of the study

To assess the feasibility of partial HPV genotyping and methylation analysis of CADM1, MAL, and miR124-2 genes as triage tests in assaying self-collected cervical samples positive for high-risk HPV on primary screening, and to review the literature regarding host cellular gene methylation analysis of self-collected cervical samples.

Material and methods

Women residing in North-East Italy who had failed to respond to the invitation to participate in an organized population-based program were invited to provide a self-sample. Their stored baseline self-collected and follow-up clinician-collected cervical samples were included in the study. DNA was extracted from HPV-positive Qiagen’s Hybrid Capture 2, HC2 samples. Partial genotyping with separate detection of HPV types 16 and 18 was performed with a hybrid capture-based method and a quantitative PCR assay. Methylation was assayed with a quantitative methylation-specific PCR.

Results

High-risk HPV infection was detected in 48% of baseline and 71% of follow-up HC2-positive samples. Methylation was demonstrated respectively in 15% and 23.5% of baseline and follow-up samples and chiefly involved a single gene miR124-2. Invalid quantitative PCR results were recorded in 5% of self-collected samples. The specificity of miR124-1, MAL, and CADM1 methylation was 84%, 94%, and 98%, respectively, and the specificity of the three markers combined was 84%. Sensitivity was not estimated due to the lack of CIN2+ samples. The systematic review showed that different methylation assays yield different accuracy values.

Conclusion

Self-collected samples are suitable for methylation assays included in reflex triage testing. The reproducibility and accuracy of the methylation tests described in the literature should be improved.



Author: Annarosa Del Mistro, Helena Frayle, Martina Rizzi, Gianpiero Fantin, Antonio Ferro, Paolo Matteo Angeletti, Paolo Giorgi Rossi, E

Source: http://plos.srce.hr/



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