96 Chapter 4 with the severity of underlying cervical disease (Kruskal-Wallis omnibus test, both p values < 0.0001). A significant increase in methylation levels was observed for both ASCL1 and LHX8 in self-collected samples from women with CIN3+ compared to self-collected samples from control women (Mann-Whitney U test, both p values < 0.0001). The ability of the individual markers and their combination to distinguish CIN3+ in hrHPVpositive self-collected samples is reported in Table 4.1. The ASCL1/LHX8 marker panel demonstrated a CIN3+ sensitivity of 73.3% (95% CI 63.9-82.6%) with a corresponding specificity of 61.1% (95% CI 56.9-65.4%). The CIN3+ sensitivity and specificity did not change with age for both the individual markers and the marker panel (p value = 0.162 and 0.377 for ASCL1 and LHX8, respectively, and p value = 0.147 for the marker panel ASCL1/ LHX8). ASCL1/LHX8 marker panel outcome in relation to HPV genotype is shown in Figure 4.3. Table 4.1 also reports the performance characteristics of HPV16/18 genotyping, HPV16/18 genotyping with ASCL1/LHX8 methylation analysis, and cytology triage. The relative sensitivity of the ASCL1/LHX8 marker panel versus HPV16/18 genotyping for CIN3+ was 0.98 (95% CI 0.82-1.17) and the relative specificity 0.92 (95% CI 0.83-1.01). The combination of HPV16/18 genotyping with ASCL1/LHX8 methylation analysis showed a ASCL1 2x NILM n = 293 0 50 100 Relative frequency LHX8 0 50 100 Relative frequency High methylation level Low methylation level Cancer n = 1 no CIN n = 71 CIN1 n = 98 CIN2 n = 45 CIN3 n = 85 Figure 4.2 Stacked histograms showing relative frequency of methylation levels of ASCL1 and LHX8 per disease category. Methylation levels were categorised in sextiles. Abbreviations: CIN, cervical intraepithelial neoplasia; 2x NILM, two consecutive normal cytology results; n, number of
RkJQdWJsaXNoZXIy MjY0ODMw