Diagnostic accuracy of the droplet digital PCR POLE mutation test in endometrial cancer: comparison with Sanger sequencing and NGS

OBJECTIVE: To evaluate the diagnostic accuracy of droplet digital polymerase chain reaction (ddPCR) for detecting polymerase epsilon (POLE) mutations in endometrial cancer (EC), comparing it with traditional Sanger sequencing and next-generation sequencing (NGS). METHODS: Tumor DNA from 132 patients with EC was analyzed initially using both ddPCR and Sanger sequencing. Discordant cases (n=20) underwent further analysis using NGS as the reference standard. This study compared the sensitivity, specificity, and overall concordance of ddPCR against Sanger sequencing results, validated by NGS. RESULTS: Among the 20 discrepant samples, ddPCR detected POLE mutations in 15 cases (75%). All ddPCR results matched exactly with NGS findings, including the detection of 11 low-frequency mutations missed by Sanger sequencing. ddPCR also corrected 2 false-positive results initially identified by Sanger sequencing. No false negatives were detected by ddPCR. CONCLUSION: ddPCR demonstrated superior sensitivity and specificity in detecting POLE mutations compared to Sanger sequencing and showed perfect agreement with NGS. With its rapid turnaround, simplicity, and cost-effectiveness, ddPCR is highly suitable for routine clinical use, potentially enhancing patient management and outcomes in EC.