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Performance and Prognostic Relevance of Lymph Node Assessment by One-Step Nucleic Acid Amplification Assay in Rectal Cancer: A Multicenter Study
by
, , , , , , , , , and
Qing Liu
1,2,†
,
Sandra Lopez-Prades
1,3,†
,
Karmele Saez de Gordoa
1,3
,
Maite Rodrigo-Calvo
1,3,
Mireia Garcia
3,
Juan Ruiz Martin
4,
Angel Romo
4,
Ignacio Pinilla
5,
Jordi Tarragona
6,
Begoña Otero Alen
7,
Jordi Camps
1,8,9,
Ivan Archilla
1,3,* and
Miriam Cuatrecasas
1,2,3,8,*
1
August Pi i Sunyer Biomedical Research Institute (IDIBAPS), 08036 Barcelona, Spain
2
Faculty of Medicine and Health Sciences, University of Barcelona, 08036 Barcelona, Spain
3
Pathology Department, Hospital Clinic Barcelona, 08036 Barcelona, Spain
4
Pathology Department, Hospital Universitario de Toledo, 45004 Toledo, Spain
5
Pathology Department, Hospital Universitario Severo Ochoa, 28911 Madrid, Spain
6
Pathology Department, Hospital Universitari Arnau de Vilanova, 25198 Lleida, Spain
7
Pathology Department, University Hospital Complex A Coruña (CHUAC), 15006 A Coruña, Spain
8
Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), 28029 Madrid, Spain
9
Cell Biology and Medical Genetics Unit, Department of Cell Biology, Physiology and Immunology, Faculty of Medicine, Autonomous University of Barcelona (UAB), 08193 Bellaterra, Spain
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Cancers 2025, 17(13), 2141; https://doi.org/10.3390/cancers17132141 (registering DOI)
Submission received: 2 June 2025
/
Revised: 19 June 2025
/
Accepted: 23 June 2025
/
Published: 25 June 2025
(This article belongs to the Section Cancer Pathophysiology)
Simple Summary
The presence of micrometastases in lymph nodes (LNs) has been associated with unfavorable prognosis in patients with colorectal cancer. A fraction of patients with stage III disease may be under-staged due to the limited sensitivity of conventional hematoxylin and eosin (H&E) analysis in detecting lymph node metastases (LNM). The One-Step Nucleic Acid Amplification (OSNA) assay has demonstrated superior performance in detecting LNM and holds prognostic significance. We aimed to assess the performance of the OSNA assay in detecting LNM and its prognostic value in rectal cancer (RC) patients. LNs were analyzed by both standard H&E and the OSNA assay. We concluded that the OSNA assay is highly sensitive for detecting LNM in RC and allows identification of a subset of RC patients with worse cancer-specific survival and recurrence-free survival who might benefit from adjuvant treatment or intensive surveillance.
Abstract
Background/Objectives: Lymph node metastases (LNM) undetected by standard hematoxylin and eosin (H&E) have been associated with unfavorable prognosis in colorectal cancer. The One-Step Nucleic Acid Amplification (OSNA) assay has demonstrated superior sensitivity in detecting LNM compared to H&E. We aimed to assess the performance of OSNA in detecting LNM, as well as its prognostic value in rectal cancer (RC) patients. Methods: Lymph nodes (LNs) of patients from 15 centers were analyzed by both H&E and OSNA. The total tumor load (TTL) was defined as the sum of cytokeratin 19 mRNA copies/µL in all LNs from a surgical specimen, using a threshold of 250 copies/μL for OSNA positivity. Cox proportional hazard regression was used to assess the effect of TTL ≥ 250 or 6000 copies/μL on cancer-specific survival (CSS) and recurrence-free survival (RFS), with Firth’s method applied to account for low event rate. Results: A total of 97 RC patients were included. Of these, 84 patients were eligible for survival analysis. The sensitivity and specificity of OSNA, compared to H&E, were 91.7% and 84.7%, respectively. TTL ≥ 6000 versus <6000 copies/μL was related to worse CSS and RFS. When dividing TTL into three groups: ≤250, 250–6000, and >6000 copies/μL, only TTL ≥ 6000 copies/μL was significantly associated with worse CSS and RFS. Conclusions: The OSNA assay is highly sensitive for detecting LNM in RC patients. A TTL of ≥6000 copies/μL could identify a subset of RC patients with worse CSS and RFS who might benefit from adjuvant treatment or intensive surveillance.
Keywords:
rectal carcinoma; lymph node metastases; staging; diagnosis; prognosis; OSNA
