Qilu Hospital, Shandong University, Jinan City, China
Beihua Kong , Qing Zhang , Guohong Hu , Qifeng Yang , Ruifen Dong , Xing Xie , Ding Ma , Keng Shen
Background: Epithelial ovarian cancer (EOC) remains the most lethal disease among gynecological malignancies. Prompt diagnosis is challenging because of the non-specific symptoms exhibited during the early stage of the disease. So there is an urgent need for better detection methods. Here we performed this work to build up a platform of multiplex methylation-specific PCR (MSP) assay to improve the early detection of ovarian cancer, via identifying the methylation status of cell-free serum DNA. Methods: After screening, we chose seven genes (APC, RASSF1A, CDH1, RUNX3, TFPI2, SFRP5 and OPCML) with a high frequency of methylation as candidate genes to construct the multiplex-MSP assay. When methylation of at least one of the seven genes was observed, the multiplex-MSP assay was considered positive. We performed the retrospective and screening study to verify its specificity and sensitivity in the detection of EOC. Results: The methylation status of cell-free serum DNA was examined in the preoperative serum of 202 patients, including 87 EOC cases (stage I, n=41, stage II-IV, n=46), 53 benign ovarian tumors and 62 healthy controls. As expected, multiplex MSP assay achieved a sensitivity of 85.3% and a specificity of 90.5% in stage I EOC, strikingly higher than that of single CA125, producing a sensitivity of 56.1% at 64.15% specificity [p=0.0036](Table). Conclusions: Multiplex MSP assay analyzing the methylation status of cell-free serum DNA is a suitable and reliable approach to improve the early detection of ovarian cancer, potentially benefiting a broad range of applications in clinical oncology.
Retrospective study |
Screening study |
||||
---|---|---|---|---|---|
CA125 | Multiplex-MSP | CA125 | Multiplex-MSP | ||
Specificity | All | 64.15% | 90.57% | 72.41% | 82.76% |
Sensitivity | All | 73.56% | 89.66% | 82.05% | 92.31% |
Early stage | 56.10% | 85.37% | 50.00% | 83.33% | |
Advanced stage | 89.13% | 93.48% | 84.85% | 93.94% | |
ROC-AUC (95% CI) |
All | 0.8090 (0.7376-0.8805) |
0.9126 ( 0.8643-0.9609) |
0.8837 (0.8017-0.9658) |
0.8988 (0.8192-0.9783) |
Early stage | 0.6986 (0.5904-0.8068) |
0.8916 (0.8258-0.9574) |
0.8103 (0.5984-1.022) |
0.8218 (0.6375-1.006) |
|
Advanced stage | 0.9075 (0.8461-0.9689) |
0.9313 (0.8788-0.9838) |
0.8970 1(0.8146-0.9796) |
0.9127 (0.8364-0.9891) |
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