• CK19
• OSNA
• breast cancer
• meta-analysis
• metastasis
• sentinel lymph nodes

• CK19
• OSNA
• meta-analysis
• metastasis
• sentinel lymph nodes

• biophotonics
• machine learning
• microscopy
• cancer

• Humans
• Lymphatic Metastasis/diagnostic imaging
• Alprostadil
• Neural Networks, Computer
• Microscopy, Fluorescence

Laparoscopic and endoscopic cooperative surgery is a safe, organ-sparing surgery that achieves full-thickness resection with adequate margins. Recent studies have demonstrated the safety and efficacy of these procedures. However, these techniques are limited by the exposure of the tumor and mucosa to the peritoneal cavity, which could lead to viable cancer cell seeding and the spillage of gastric juice or enteric liquids into the peritoneal cavity. Non-exposed endoscopic wall-inversion surgery (NEWS) is highly accurate in determining the resection margins to prevent intraperitoneal contamination because the tumor is inverted into the visceral lumen instead of the peritoneal cavity. Accurate intraoperative assessment of the nodal status could allow stratification of the extent of resection. One-step nucleic acid amplification (OSNA) can provide a rapid method of evaluating nodal tissue, whilst near-infrared laparoscopy together with indocyanine green can identify relevant nodal tissue intraoperatively.

To determine the safety and feasibility of NEWS in early gastric and colon cancers and of adding rapid intraoperative lymph node (LN) assessment with OSNA.

The patient-based experiential portion of our investigations was conducted at the General and Oncological Surgery Unit of the St. Giuseppe Moscati Hospital (Avellino, Italy). Patients with early-stage gastric or colon cancer (diagnosed via endoscopy, endoscopic ultrasound, and computed tomography) were included. All lesions were treated by NEWS procedure with intraoperative OSNA assay between January 2022 and October 2022. LNs were examined intraoperatively with OSNA and postoperatively with conventional histology. We analyzed patient demographics, lesion features, histopathological diagnoses, R0 resection (negative margins) status, adverse events, and follow-up results. Data were collected prospectively and analyzed retrospectively.

A total of 10 patients (5 males and 5 females) with an average age of 70.4 ± 4.5 years (range: 62-78 years) were enrolled in this study. Five patients were diagnosed with gastric cancer. The remaining 5 patients were diagnosed with early-stage colon cancer. The mean tumor diameter was 23.8 ± 11.6 mm (range: 15-36 mm). The NEWS procedure was successful in all cases. The mean procedure time was 111.5 ± 10.7 min (range: 80-145 min). The OSNA assay revealed no LN metastases in any patients. Histologically complete resection (R0) was achieved in 9 patients (90.0%). There was no recurrence during the follow-up period.

NEWS combined with sentinel LN biopsy and OSNA assay is an effective and safe technique for the removal of selected early gastric and colon cancers in which it is not possible to adopt conventional endoscopic resection techniques. This procedure allows clinicians to acquire additional information on the LN status intraoperatively.

• Laparoscopic and endoscopic cooperative surgery
• Non-exposed endoscopic wall inversion surgery
• Early gastric cancer
• Early colorectal cancer
• Sentinel lymph node
• One-step nucleic acid amplification
• Endoscopic full-thickness resection

• Aged
• Female
• Humans
• Male
• Colonic Neoplasms/genetics
• Colonic Neoplasms/pathology
• Colonic Neoplasms/surgery
• Laparoscopy/adverse effects
• Laparoscopy/methods
• Lymph Nodes/pathology
• Lymphatic Metastasis/pathology
• Retrospective Studies
• Sentinel Lymph Node Biopsy/methods
• Stomach Neoplasms/genetics
• Stomach Neoplasms/surgery
• Stomach Neoplasms/pathology
• Organ Sparing Treatments
• Gastrointestinal Neoplasms/genetics
• Gastrointestinal Neoplasms/pathology
• Gastrointestinal Neoplasms/surgery
• Esophageal Neoplasms/genetics
• Esophageal Neoplasms/pathology
• Esophageal Neoplasms/surgery
• Nucleic Acid Amplification Techniques

• Cervical cancer
• Endometrial cancer
• One-step nucleic acid amplification
• Sentinel lymph node

• Humans
• Female
• Lymphatic Metastasis/pathology
• Prospective Studies
• Nucleic Acid Amplification Techniques/methods
• Endometrial Neoplasms/pathology
• Nucleic Acids
• Lymph Nodes/pathology
• Sentinel Lymph Node Biopsy
• Keratin-19/genetics
• Breast Neoplasms/pathology

• PCR
• TERTp
• intraoperative
• meningioma
• molecular diagnosis
• surgery

• OSNA assay
• cytokeratin 19
• lung cancer
• lymph nodes
• metastasis

• OSNA
• biomarkers
• breast neoplasms
• immune system
• immunotherapy
• lymph nodes
• metastases

• POCI-01-0145-FEDER-022184 GenomePT-National Laboratory for Genome Sequencing and Analysis
• CENTRO-08-5864-FSE-000039 (PEP IN1194) 07-2021 Central Region Training Project for Personalized/Precision Medicine, with a genomic basis

• Breast cancer
• Breast-conserving surgery
• Incidental irradiation
• Nodal irradiation
• OSNA
• Sentinel lymph node

• Humans
• Female
• Lymphatic Metastasis/pathology
• Neoplasm Recurrence, Local/pathology
• Lymph Nodes/pathology
• Sentinel Lymph Node Biopsy
• Axilla/pathology
• Lymph Node Excision
• Breast Neoplasms/genetics
• Breast Neoplasms/radiotherapy
• Breast Neoplasms/pathology

• CEA
• OSNA
• colorectal cancer
• micrometastasis

• Keywords sentinel lymph node biopsy
• OSNA
• breast cancer
• frozen section
• survival

One-step nucleic acid amplification (OSNA) is a rapid intraoperative molecular detection technique for sentinel node assessment via the quantitative measurement of target cytokeratin 19 (CK19) mRNA to determine the presence of metastasis. It has been validated in breast cancer but its application in lung cancer has not been adequately investigated. 214 LNs from 105 patients with 100 primary lung cancers, 2 occult primary lung tumors, and 3 metastatic lung tumors, who underwent surgical lung resection with LN dissection between February 2018 and January 2020, were assessed. Resected LNs were divided into two parts: one was snap-frozen for OSNA and the other underwent rapidly frozen histological examination. Intraoperatively collected LNs were evaluated by OSNA using loop-mediated isothermal amplification and compared with intraoperative pathological diagnosis as a control. Among 214 LNs, 14 were detected as positive by OSNA, and 11 were positive by both OSNA and intraoperative pathological diagnosis. The sensitivity and specificity of OSNA was 84.6% and 98.5%, respectively. The results of 5 of 214 LNs were discordant, and the remainder all matched (11 positive and 198 negative) with a concordance rate of 97.7%. Although the analysis of public mRNA expression data from cBioPortal showed that CK19 expression varies greatly depending on the cancer type and histological subtype, the results of the five cases, except for primary lung cancer, were consistent. OSNA provides sufficient diagnostic accuracy and speed and can be applied to the intraoperative diagnosis of LN metastasis for non-small cell lung cancer.

The prognosis of non-small cell lung cancer greatly depends on the presence of lymph node metastasis, which limits the need for surgery and adjuvant therapy for advanced cancer. One-step nucleic acid amplification of cytokeratin19 (CK19) mRNA was used to detect lymph node metastasis. Automated Gene Amplification Detector RD-200 and the LYNOAMP CK19 gene amplification reagent as components of the new one-step nucleic acid amplification system, which has increased gene amplification efficiency by improving the reagent composition, have shorter preprocessing and measurement times than conventional systems. We aimed to compare the clinical performance of the new system with that of histopathology and the conventional system.

199 lymph nodes from 58 non-small cell lung cancer patients who underwent lymph node dissection were examined intraoperatively using the new system, conventional system, and histopathology.

Lymph node metastasis was diagnosed in 32, 42, and 44 patients using histopathological analysis, the new system, and the conventional system, respectively. Compared with histopathological analysis, the concordance rate, sensitivity, specificity, positive predictive value, and negative predictive value of the new system were 92.0%, 90.6%, 92.2%, 69.0%, and 98.1%, respectively, and compared with the conventional system, the values were 95.0%, 86.4%, 97.4%, 90.5%, and 96.2%, respectively.

The clinical performance of the new one-step nucleic acid amplification system in detecting lymph node metastasis of lung cancer is comparable to that of histopathology and the conventional system; its performance was sufficient for determining the appropriate clinical treatment. The new rapid system can be effectively utilized during lung cancer treatment intraoperatively and postoperatively.

• breast cancer
• cytokeratin 19
• multicenter study
• one-step nucleic acid amplification (OSNA) assay
• prediction model
• sentinel lymph node
• total tumor load

Although lobectomy is considered the standard surgery for any non‐small cell lung cancer (NSCLC), recent evidence indicates that for early NSCLCs segmentectomy may be equally effective. For segmentectomy to be oncologically safe, however, adequate intraoperative lymph node staging is essential. The aim of this study was to compare the results of a new rapid‐IHC system to the HE analysis for intraoperative nodal diagnosis in lung cancer patients considered for segmentectomy.

This retrospective study analyzed the pathological reports from NSCLC resections over a six‐year period between 2014 and 2020. Using a new device for rapid‐IHC, we applied a high‐voltage, low‐frequency alternating current (AC) field, which mixes the antipancytokeratin antibody as the voltage is switched on/off. Rapid‐IHC can provide a nodal diagnosis within 20 minutes.

Frozen sections from 106 resected lymph nodes from 70 patients were intraoperatively evaluated for metastasis. Of those, five nodes were deemed positive based on both HE staining and rapid‐IHC. In addition, rapid‐IHC alone detected isolated tumor cells in one hilar lymph node. Three cStage IA patients with nodal metastasis detected with HE staining and rapid‐IHC received complete lobectomies. Five‐year relapse‐free survival and overall survival among patients receiving segmentectomy with rapid‐IHC were 88.77% and 88.79%, respectively.

Rapid‐IHC driven by AC mixing is simple, highly accurate, and preserves nodal tissue for subsequent tests. This system can be used effectively for intraoperative nodal diagnosis.

Rapid immunohistochemistry based on alternating‐current field mixing (completed within 20 minutes) is simple and highly accurate. This system will assist clinicians when making intraoperative diagnoses of lymph node metastasis and deciding upon the appropriate surgical procedure in segmentectomy for lung cancer.

Rapid immunohistochemistry driven by alternating‐current field mixing (completed within 20 minutes intraoperatively) is simple, highly accurate, and preserves lymph node tissue for subsequent pathological examination, including molecular assessments.

Segmentectomy for lung cancer is oncologically safe, but only when there is adequate intraoperative node staging. Rapid immunohistochemistry will assist clinicians when making intraoperative nodal diagnoses.

• Intraoperative diagnosis
• lung cancer
• lymph node metastasis
• rapid immunohistochemistry
• segmentectomy

• Breast cancer
• Invasive carcinoma
• Metastases
• OSNA
• Sentinel lymph node procedure

• Adult
• Aged
• Aged, 80 and over
• Breast Neoplasms/pathology
• Cohort Studies
• Female
• Humans
• Medical Overuse
• Middle Aged
• Neoplasm Micrometastasis/pathology
• Netherlands
• Nucleic Acid Amplification Techniques/methods
• Nucleic Acid Amplification Techniques/statistics & numerical data
• Prospective Studies
• Retrospective Studies
• Sentinel Lymph Node/pathology
• Sentinel Lymph Node Biopsy/methods

• Breast cancer
• Molecular axillary staging
• OSNA
• Prognosis

• Endosalpingiosis
• Gynecological cancer
• Isolated tumor cells
• Liquid-based cytology
• Sentinel lymph node

• Gastrectomy
• Gastric cancer
• Histopathology
• OSNA assay
• Sentinel node mapping

The aim of the study is to construct an intraoperative nomogram for the prediction of non-sentinel lymph node (NSLN) metastasis based on the one-step nucleic acid amplification assay in breast cancer patients.

A total of 552 patients were enrolled in the training study and 1090 patients were enrolled in the validation study. The nomogram was constructed based on the molecular assay with logistic multivariate regression analysis in the training study and was validated in the validation study.

A novel nomogram model was constructed with the total tumor load, the clinical primary tumor size, the number of positive and negative sentinel lymph nodes. The area under the receiver operating characteristic curve (AUC) of the model was 0.842. The AUC of the model which was sensitive to discern the patients with the stage of pN1 and ≥pN2 was 0.861.

The nomogram model will help to guide the axillary management intraoperatively and precisely confirm the target region of radiotherapy postoperatively.

• breast neoplasm
• molecular diagnostic techniques
• nomograms
• sentinel lymph node biopsy

• Intraoperative
• Keratin 19
• LN status
• Lung cancer
• Metastasis
• OSNA assay

• Fluorescence imaging
• Gastric cancer
• Indocyanine green
• Sentinel lymph node
• Surgical pathology

• Apoptosis
• Circulating tumor DNA
• PIK3CA mutation
• Sentinel lymph node

• Breast cancer
• DCIS
• Lymph node metastasis
• OSNA
• Parallel progression model
• Survival

• Blood/microbiology
• Drug Resistance, Microbial
• Humans
• Monitoring, Intraoperative/instrumentation
• Monitoring, Intraoperative/methods
• Point-of-Care Systems
• Single-Cell Analysis
• Skin/diagnostic imaging
• Software
• Spectroscopy, Fourier Transform Infrared/methods
• Spectrum Analysis, Raman/methods
• Urine/microbiology

• R01 CA197516 NCI NIH HHS
• R01 EB009745 NIBIB NIH HHS
• R01 GM117594 NIGMS NIH HHS

One-step nucleic acid amplification (OSNA) for cytokeratin 19 messenger RNA is an intraoperative diagnostic procedure for the detection of lymph node metastasis.

This study aimed to construct intraoperative nomograms using OSNA for the prediction of non-sentinel lymph node (NSLN) metastasis and four or more axillary lymph node (ALN) metastases.

Of the 4736 breast cancer patients (T1-3, N0) who underwent sentinel lymph node (SLN) biopsy and had SLNs examined intraoperatively with OSNA, 623 with SLN metastasis treated with completion ALN dissection (cALND) were retrospectively analyzed, and were randomly divided into training (n = 312) and validation (n = 311) sets.

Of the clinicopathological parameters available preoperatively and intraoperatively, the multivariate analysis of the training set revealed that clinical tumor size and total tumor load (TTL) determined by OSNA were significantly associated with NSLN metastasis, and that clinical tumor size, number of macrometastatic SLNs, and TTL were significantly associated with four or more ALN metastases. Nomograms for NSLN metastasis and four or more ALN metastases were constructed using these parameters, and their area under the receiver operating characteristic curve (AUC) of the validation set were both 0.70, with a diagnostic accuracy similar to that of previously reported postoperative nomograms.

We constructed intraoperative nomograms using OSNA for the prediction of NSLN metastasis and four or more ALN metastases. These nomograms are as accurate as the conventional postoperative nomograms and might be helpful for decision making regarding the indication for cALND or the choice of adjuvant chemotherapeutic regimens and radiation field.

The online version of this article (10.1245/s10434-018-6633-0) contains supplementary material, which is available to authorized users.

• breast cancer
• intraoperative assessment
• mastectomy
• one-stop nucleic acid amplification
• sentinel lymph node

• Breast cancer
• CK-IHC
• Intraoperative diagnosis
• OSNA
• SLN

The one-step nucleic acid amplification (OSNA) assay is a novel molecular method that can detect metastasis in a whole lymph node based on cytokeratin 19 mRNA copy number. This cohort study aimed to establish an OSNA-based nodal staging (pN(mol)) classification for breast cancer.

The cohort consisted of 1039 breast cancer patients who underwent sentinel node (SN) biopsy using the OSNA assay. Cutoff value of the SN tumour burden stratifying distant disease-free survival (DDFS) was determined, and predictive factors for DDFS and breast cancer-specific survival (BCSS) were investigated. pN(mol) classification of the SN status was defined as: pN0(mol)(sn), SN negative; pN1mi(mol)(sn), SN positive and tumour burden <cutoff-value; and pN1(mol)(sn), tumour burden ⩾cutoff-value. Median follow-up time; 68.3 months.

Cutoff value of the SN tumour burden was 2810 copies per μl. Of the 1039 patients, 798, 95, and 146 had pN0(mol)(sn), pN1mi(mol)(sn), and pN1(mol)(sn) status, respectively. Five-year DDFS and BCSS rates were lower for pN1(mol)(sn) patients than for pN1mi(mol)(sn) patients (87.7% vs 98.8%, P=0.001 and 93.1% vs 98.8%, P=0.044, respectively). Multivariate analyses revealed the pN(mol) classification was most significant predictor for DDFS and BCSS.

The molecular-based pN classification determines the prognosis of breast cancer patients and could guide therapeutic decision making.

Since 2007, one-step nucleic acid amplification (OSNA) has been used as a diagnostic system for sentinel lymph node (SLN) examination in patients with breast cancer. This study aimed to define a new clinical cut-off of CK19 mRNA copy number based on the calculation of the risk that an axillary lymph node dissection (ALND) will be positive. We analyzed 1529 SLNs from 1140 patients with the OSNA assay and 318 patients with positive SLNs for micrometastasis (250 copies) and macrometastasis (5000 copies) underwent ALND. Axillary non–SLNs were routinely examined. ROC curves and Youden’s index were performed in order to identify a new cut-off value. Logistic regression models were performed in order to compare OSNA categorical variables created on the basis of our and traditional cut-off to better identify patients who really need an axillary dissection. 69% and 31% of OSNA positive patients had a negative and positive ALND, respectively. ROC analysis identified a cut-off of 2150 CK19 mRNA copies with 95% sensitivity and 51% specificity. Positive and negative predictive values of this new cut-off were 47% and 96%, respectively. Logistic regression models indicated that the cut-off of 2150 copies better discriminates patients with node negative or positive in comparison with the conventional OSNA cut-off (p<0.0001). This cut-off identifies false positive and false negative cases and true-positive and true negative cases very efficiently, and therefore better identifies which patients really need an ALND and which patients can avoid one. This is why we suggest that the negative cut-off should be raised from 250 to 2150. Furthermore, we propose that for patients with a copy number that ranges between 2150 and 5000, there should be a multidisciplinary discussion concerning the clinical and bio-morphological features of primary breast cancer before any decision is taken on whether to perform an ALND or not.

The sentinel lymph node (SLN) biopsy is a highly accurate predictor of overall axillary nodal status in early breast cancer patients. There is however, still a debate on which patients with a positive SLN can benefit from axillary lymph node dissection (ALND). Numerous studies have been designed to identify variables that are predictive of non-SLN metastasis to avoid a complete ALND. The aim of this study was to determine whether the pre-treatment neutrophil-lymphocyte ratio (NLR) can be a predictive factor of non-SLN metastasis in early breast cancer patients.

The records of 214 consecutive patients with cT1-3N0 invasive breast cancer who had undergone intraoperative SLN evaluation at Songklanagarind Hospital between the 1st of March 2011 and the 30th of May 2016 were examined. Data on patient demographics, tumor variables and NLR were collected and factors for non-SLN metastasis were analyzed using multivariate logistic regression. The power of the NLR was quantified with receiver operating characteristics (ROC) curves as measured by the areas under curves (AUC).

Multivariate analysis established presence of lymphovascular invasion (OR 8.4, 95%CI 2.3-31.3, p=0.002), macrometastasis (OR 6.6, 95%CI 1.8-24.7, p=0.005), and NLR (OR 2.3, 95%CI 1.1-4.8, p=0.033) as predictive factors of non-SLN metastasis with statistical significance. The AUC for NLR was 0.7 (95%CI 0.6-0.8) with an optimal cut-off of 2.6 giving a sensitivity of 62%, a specificity of 83.8%, a positive predictive value of 77.3% and a negative predictive value of 70.5%.

Pre-treatment NLR is a useful diagnostic aid for predicting additional non-SLN metastasis.

• Neutrophil-lymphocyte ratio
• NLR
• breast cancer
• non sentinel lymph node

Pathologic lymph node staging is becoming a deficient method in the demanding molecular era. Nevertheless, the use of more sensitive molecular analysis for nodal staging is hampered by its high costs and extensive time requirements. Our aim is to take a step forward in colon cancer (CC) lymph node (LN) pathology diagnosis by proposing a feasible and efficient molecular method in routine practice using reverse transcription loop-mediated isothermal amplification (RT-LAMP).

Molecular detection of tumor cytokeratin 19 (CK19) mRNA with RT-LAMP was performed in 3206 LNs from 188 CC patients using two methods: individual analysis of 1449 LNs from 102 patients (individual cohort), and pooled LN analysis of 1757 LNs from 86 patients (pooling cohort). A median of 13 LNs (IQR 10–18) per patient were harvested in the individual cohort, and 18 LNs (IQR 13–25) per patient in the pooling cohort (p ≤ 0.001). The median of molecular assays performed in the pooling cohort was 2 per patient (IQR 1–3), saving a median of 16 assays/patient. The number of molecular assays performed in the individual cohort was 13 (IQR 10–18), corresponding to the number of LNs to be analyzed. The sensitivity and specificity of the pooling method for LN involvement (assessed by hematoxylin and eosin) were 88.9% (95% CI 56.5–98.0) and 79.2% (95% CI 68.9–86.8), respectively; concordance, 80.2%; PPV, 33.3%; NPV, 98.4%. The individual method had 100% sensitivity (95% CI 72.2–100), 44.6% specificity (95% CI 34.8–54.7), 50% concordance, 16.4% PPV, and 100% NPV. The amount of tumor burden detected in all LNs of a case, or total tumor load (TTL) was similar in both cohorts (p = 0.228).

LN pooling makes it possible to analyze a high number of LNs from surgical colectomies with few molecular tests per patient. This approach enables a feasible means to integrate LN molecular analysis from CC specimens into pathology diagnosis and provides a more accurate LN pathological staging with potential prognostic implications.

• Colon cancer
• Lymph node
• Molecular
• OSNA
• Pooling
• Staging
• Total tumor load

• breast cancer
• cytokeratin 19
• intraoperative molecular study
• sentinel lymph node

Cervical lymph node metastasis is an important prognostic factor in oral squamous cell carcinoma (OSCC), but its accurate assessment after sentinel node biopsy or neck dissection is often limited to the histopathological examination of only one or two sections. Previous our study showed the usefulness of the reverse transcription loop-mediated isothermal amplification (RT-LAMP) targeting keratin 19 (KRT19) mRNA for the genetic detection of lymph node metastasis, but the sensitivity was insufficient. Here, we have attempted to identify novel molecular markers for OSCC cells in lymph nodes. We performed microarray analysis to identify genes overexpressed in 7 metastatic lymph nodes from OSCC patients, compared to 1 normal lymph node and 5 salivary glands from non-cancer patients. We then used real-time quantitative RT-PCR (qRT-PCR) and RT-LAMP to compare the expression of these genes in newly resected metastatic and normal lymph nodes. Of 4 genes identified by microarray analysis, annexin A8 (ANXA8) and desmoglein 3 mRNA were detected by qRT-PCR in metastatic lymph nodes but not in normal lymph nodes. Furthermore, ANXA8 mRNA expression was detected in all KRT19-negative metastatic lymph nodes. Both KRT19 and ANXA8 mRNA may be useful markers for detecting lymph node metastases in OSCC patients.

• annexin A8 (ANXA8)
• keratin 19 (KRT19)
• lymph node metastasis
• oral squamous cell carcinoma (OSCC)
• reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Stage I–II (pN0) colorectal cancer patients are surgically treated although up to 25 % will eventually die from disease recurrence. Lymph node (LN) status is an independent prognostic factor in colorectal cancer (CRC), and molecular tumour detection in LN of early-stage CRC patients is associated with an increased risk of disease recurrence and poor survival. This prospective multicentre study aimed to determine the relationship between LN molecular tumour burden and conventional high-risk factors in stage I–II colon cancer patients. A total of 1940 LN from 149 pathologically assessed pN0 colon cancer patients were analysed for the amount of tumour cytokeratin 19 (CK19) messenger RNA (mRNA) with the quantitative reverse transcription loop-mediated isothermal amplification molecular assay One-Step Nucleic Acid Amplification. Patient’s total tumour load (TTL) resulted from the sum of all CK19 mRNA tumour copies/μL of each positive LN from the colectomy specimen. A median of 15 LN were procured per case (IQR 12;20). Molecular positivity correlated with high-grade (p < 0.01), mucinous/signet ring type (p = 0.017), male gender (p = 0.02), number of collected LN (p = 0.012) and total LN weight per case (p < 0.01). The TTL was related to pT stage (p = 0.01) and tumour size (p < 0.01) in low-grade tumours. Multivariate logistic regression showed independent correlation of molecular positivity with gender, tumour grade and number of fresh LN [AUC = 0.71 (95 % CI = 0.62–0.79)]. Our results show that lymph node CK19 mRNA detection correlates with classical high-risk factors in stage I–II colon cancer patients. Total tumour load is a quantitative and objective measure that may help to better stage early colon cancer patients.

• Colorectal neoplasms
• Cytokeratin 19
• Lymph nodes
• Molecular pathology
• Neoplasm staging

• Aged
• Colonic Neoplasms/diagnosis
• Colonic Neoplasms/pathology
• Female
• Humans
• Lymph Nodes/pathology
• Lymphatic Metastasis
• Male
• Middle Aged
• Neoplasm Recurrence, Local/diagnosis
• Neoplasm Recurrence, Local/pathology
• Neoplasm Staging/methods
• Prospective Studies
• Risk Factors
• Tumor Burden

• Fundación Científica de la Asociación Española Contra el Cáncer
• Ministerio de Economía y Competitividad
• Agència de Gestió d’Ajuts Universitaris i de Recerca

• Histological examination
• LN status
• Lymph node metastasis
• Non-small cell lung cancer (NSCLC)
• One-step nucleic acid amplification (OSNA)