Application of Multiplex Nested Methylated Specific PCR in Early Diagnosis of Epithelial Ovarian Cancer

Abstract

<br/><b>Objective</b>: To explore the application of multiplex nested methylated specific polymerase chain reaction (PCR)in the early diagnosis of epithelial ovarian carcinoma (EOC). Materials and <br/><b>Methods</b>: Serum and fresh tissuesamples were collected from 114 EOC patients. RUNX3, TFPI2 and OPCML served as target genes. Methylationlevels of tissues were assessed by multiplex nested methylated specific PCR, the results being compared withthose for carcinoma antigen 125 (CA125). <br/><b>Results</b>: The serum free deoxyribose nucleic acid (DNA) methylationspectrum of EOC patients was completely contained in the DNA spectrum of cancer tissues, providing an accuratereflection of tumor DNA methylation conditions. Serum levels of CA125 and free DNA methylation in the EOCgroup were evidently higher than those in benign lesion and control groups (p<0.05). Patients with early EOC hadmarkedly lower serum CA125 than those with advanced EOC (p<0.05), but there was no significant differencein free DNA methylation (p>0.05). The sensitivity, specificity and positive predicative value (PPV) of multiplexnested methylated specific PCR were significantly higher for detection of all patients and those with early EOCthan those for CA125 (p<0.05). In the detection of patients with advanced EOC, the PPV of CA125 detectionwas obviously lower than that of multiplex nested methylated specific PCR (p>0.05), but there was no significantdifference in sensitivity (p>0.05). <br/><b>Conclusions</b>: Serum free DNA methylation can be used as a biological markerfor EOC and multiplex nested methylated specific PCR should be considered for early diagnosis since it canaccurately determine tumor methylation conditions.