Hesselink A T, Heideman D A M, Steenbergen R D M, Gök M, van Kemenade F J, Wilting S M, Berkhof J, Meijer C J L M, Snijders P J F
Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands.
Int J Cancer. 2014 Aug 15;135(4):880-6. doi: 10.1002/ijc.28723. Epub 2014 Jan 28.
Methylation markers were studied for their suitability to triage human papillomavirus (HPV)-positive women by testing self-collected cervico-vaginal lavage specimens. For this purpose, we analyzed 355 hrHPV-positive self-collected specimens with three methylation markers, that is, CADM1-m18, MAL-m1 and miR-124-2 by quantitative methylation-specific PCR. The areas under the receiver-operating characteristic (ROC) curve for end-point cervical intraepithelial neoplasia grade 3 or worse (CIN3+) were 0.637 for CADM1-m18, 0.767 for MAL-m1 and 0.762 for miR-124-2. This indicates that CADM1-m18 is not suitable as single marker. By varying the thresholds of both markers in the bi-marker panels CADM1-m18/MAL-m1, CADM1-m18/miR-124-2 and MAL-m1/miR-124-2 upper and lower ROC curves were obtained, depicting the maximum and minimum CIN3+ sensitivity, respectively, at given specificity. For all these bi-marker combinations, the upper curves were similar. However, for the MAL-m1/miR-124-2 panel, the distance between upper and lower ROC curves was closest and this panel displayed the highest assay thresholds, indicating that this combination was most robust. At clinical specificities of 50 and 70%, the MAL-m1/miR-124-2 sensitivity for detection of CIN3+ ranged from 77.0 to 87.8% and from 64.9 to 71.6%, respectively. At 70% specificity thresholds no carcinomas were missed. By comparison, the CIN3+ sensitivity of HPV16/18 genotyping on the self-sampled lavage specimens was 58.1% (95%CI: 46.6-68.8) at a specificity of 87.7% (95%CI: 83.2-91.2). In conclusion, methylation analysis is a promising triage tool that in combination with HPV-DNA testing offers feasible, full molecular screening on self-collected cervico-vaginal lavage specimens.
通过检测自行采集的宫颈阴道灌洗样本，研究甲基化标志物对人乳头瘤病毒（HPV）阳性女性进行分流的适用性。为此，我们采用定量甲基化特异性PCR分析了355份高危型HPV（hrHPV）阳性的自行采集样本，检测了三种甲基化标志物，即CADM1-m18、MAL-m1和miR-124-2。终点宫颈上皮内瘤变3级及以上（CIN3+）的受试者操作特征（ROC）曲线下面积，CADM1-m18为0.637，MAL-m1为0.767，miR-124-2为0.762。这表明CADM1-m18不适合作为单一标志物。通过改变双标志物组合CADM1-m18/MAL-m1、CADM1-m18/miR-124-2和MAL-m1/miR-124-2中两种标志物的阈值，获得了上下ROC曲线，分别描绘了在给定特异性下CIN3+的最大和最小敏感性。对于所有这些双标志物组合，上曲线相似。然而，对于MAL-m1/miR-124-2组合，上下ROC曲线之间的距离最接近，且该组合显示出最高的检测阈值，表明该组合最为稳健。在临床特异性为50%和70%时，MAL-m1/miR-124-2检测CIN3+的敏感性分别为77.0%至87.8%和64.9%至71.6%。在特异性阈值为70%时，没有漏诊癌症。相比之下，自行采集的灌洗样本上HPV16/18基因分型检测CIN3+的敏感性在特异性为87.7%（95%CI：83.2-91.2）时为58.1%（95%CI：46.6-68.8）。总之，甲基化分析是一种很有前景的分流工具，与HPV-DNA检测相结合，可对自行采集的宫颈阴道灌洗样本进行可行的全分子筛查。
