2025

Szeto, Säde; Kytölä, Soili; Erkan, Erdogan Pekcan; Ahtiainen, Maarit; Mecklin, Jukka-Pekka; Kuopio, Teijo; Sallinen, Ville; Lepistö, Anna; Koskenvuo, Laura; Renkonen-Sinisalo, Laura; Anttonen, Anu; Heiskala, Kukka; Tulokas, Sanni; Mäkelä, Siru; Wirta, Erkki-Ville; Tuunanen, Tuija; Salminen, Tapio; Ristimäki, Ari; Seppälä, Toni T.;

Background and Objectives Circulating tumor DNA (ctDNA) can potentially identify rectal cancer patients benefiting from neoadjuvant and adjuvant therapy. This study compared droplet digital PCR (ddPCR) and next-generation sequencing (NGS) for ctDNA detection in localized rectal cancer before and after surgery. Methods Pre-therapy plasma and rectal tumor samples were collected from a development group (n = 41) and a validation group (n = 26). Mutations in tumor samples were identified using NGS, and ctDNA detection was performed with both ddPCR and NGS. Recurrence was assessed 1 year after surgery in the development group. Results In the development group, ddPCR detected ctDNA in 24/41 (58.5%) and NGS panel in 15/41 (36.6%; p = 0.00075) of the baseline plasma. In the validation group, 21/26 (80.8%) patients had detectable ctDNA in the pre-therapy plasma. A positive ctDNA result was associated with higher clinical tumor stage and with lymph node positivity as detected by MRI. Postoperative ddPCR did not detect ctDNA before most recurrences. Conclusions We demonstrated a practical oligomarker ctDNA test for localized rectal cancer suitable for clinical workflow, and that ddPCR detects ctNA from pre-therapy plasma at a satisfactory level in advanced rectal cancers. Detecting ctDNA with ddPCR may help to assess the local severity, but the clinical utility of this approach should be evaluated in clinical trials.