• Breast cancer
• diagnosis
• intraoperative
• lymph node
• metasin
• molecular
• one-step nucleic acid amplification (OSNA)
• sentinel node
• surgery

• Auto-fluorescence imaging
• Breast cancer
• Lymph node metastasis
• Raman spectroscopy
• Spectral imaging

• Humans
• Breast Neoplasms/surgery
• Breast Neoplasms/pathology
• Female
• Spectrum Analysis, Raman/methods
• Sentinel Lymph Node/pathology
• Sentinel Lymph Node/surgery
• Sentinel Lymph Node Biopsy/methods
• Middle Aged
• Lymphatic Metastasis/pathology
• Aged
• ROC Curve
• Sensitivity and Specificity
• Adult
• Optical Imaging/methods

• 2017NovPR1009 Breast Cancer Now

• Animals
• Mice
• Humans
• Female
• Lymphatic Metastasis/diagnostic imaging
• Lymphatic Metastasis/pathology
• Sentinel Lymph Node/diagnostic imaging
• Sentinel Lymph Node/pathology
• Breast Neoplasms/pathology
• Lymph Nodes/diagnostic imaging
• Lymph Nodes/pathology
• Neoplasm Micrometastasis/pathology
• Sentinel Lymph Node Biopsy/methods
• Neoplasm Staging
• Axilla/pathology

• National Natural Science Foundation of China (NSFC)

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

• Breast cancer
• Intra-operative assessment
• Lymph nodes
• Raman spectroscopy

• OSNA
• cytokeratin 19
• metastases
• prostate cancer
• sentinel lymph node

• 2016 Research Pool of Carl von Ossietzky University Oldenburg, Oldenburg, Germany

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

• Adenocarcinoma/genetics
• Adenocarcinoma/pathology
• Adenocarcinoma/surgery
• Biomarkers, Tumor/analysis
• Biomarkers, Tumor/genetics
• Female
• Follow-Up Studies
• Gastrectomy
• Humans
• Male
• Middle Aged
• Nucleic Acid Amplification Techniques/methods
• Prognosis
• Prospective Studies
• Sentinel Lymph Node/pathology
• Sentinel Lymph Node/surgery
• Sentinel Lymph Node Biopsy
• Stomach Neoplasms/genetics
• Stomach Neoplasms/pathology
• Stomach Neoplasms/surgery

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.

The widely practiced intra-operative methods for rapid evaluation and detection of sentinel lymph node (SLN) status include frozen section (FS) and touch imprint cytology (TIC). This study optimized the use of TIC and FS in the intra-operative detection of breast SLNs based on the Memorial Sloan Kettering Cancer Center (MSKCC) nomogram. Three hundred forty-two SLNs were removed from 79 patients. SLN metastatic probability was assessed by the MSKCC nomogram. The SLNs underwent intra-operative TIC and FS, as well as routine post-operative paraffin sections (RPSs). The relationships between TIC, FS, and SLN metastatic probability were analyzed. Overall, TIC was more sensitive than FS (92.31% vs. 76.92%), while TIC specificity was inferior to FS specificity (84.85% vs. 100%). In addition, the best cut-off value for TIC based on the MSKCC nomogram was inferior to the best FS cut-off value (22.5% vs. 34.5%). All patients with a MSKCC value <22.5% in the present study were negative based on FS and RPS, while the true-negative and false-positive rates for TIC were 92.5% and 7.5%, respectively. Thus, early breast cancer patients, based on a MSKCC value <22.5%, can safely avoid FS, but should have TIC performed intra-operatively. Patients with a MSKCC value >22.5% should have TIC and FS to determine the size of metastases, whether or not to proceed with axillary lymph node dissection, and to avoid easily missed metastases.

• Memorial Sloan Kettering Cancer Center nomogram
• early breast cancer
• frozen section
• intraoperative detection
• touch imprint cytology

To estimate the accuracy of one-step nucleic acid amplification (OSNA) assay as an intra-operative sentinel lymph node biopsy (SLNB) for sentinel lymph node (SLN) metastasis in breast cancer.

PubMed, Cochrane Library and Web of Science databases were searched by two independent reviewers to retrieve literature with per-patient analysis. The deadline was up until December 2016. A meta-analysis was performed using STATA, Meta-Disc, and Revman software. A random-effects model was used and subgroup analysis was carried out to identify possible sources of heterogeneity.

According to the inclusion criteria, 2833 patients from 12 studies were included in this meta-analysis. The pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR) and the area under the sROC curve (AUC) for detecting SLN metastasis were 0.87 (95% CI 0.81–0.91), 0.92 (95% CI 0.86–0.95), 10.65 (95% CI 6.18–18.34), 0.14 (95% CI 0.10–0.20), 75.08 (95% CI 37.77–149.22) and 0.94 (95% CI 0.91–0.95), respectively.

The present study adds the evidence that OSNA assay is an accurate molecular diagnostic tool for intra-operatively detecting SLN metastasis in breast cancer. One-step nucleic acid amplification assay might be introduced into clinical usage for replacing traditional intro-operative diagnostic methods of SLNB.

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.

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

• CMOS
• diagnostic
• integrated biosensor
• intraoperative detection
• lymph node metastasis
• multichannel detection
• nano fabrication
• silicon nanowire on a chip

• Animals
• Biosensing Techniques/instrumentation
• Biosensing Techniques/methods
• Lymph Nodes/pathology
• Mice
• Molecular Diagnostic Techniques/instrumentation
• Molecular Diagnostic Techniques/methods
• Nanowires/chemistry
• Neoplastic Cells, Circulating/pathology
• Sensitivity and Specificity
• Silicon
• Transistors, Electronic

• invasive breast cancer
• nanoparticles
• sentimag®
• sentinel lymph node biopsy
• sienna®
• superparamagnetic iron oxide (SPIO)

• Aged
• Breast Neoplasms/pathology
• Breast Neoplasms/surgery
• Carcinoma/secondary
• Carcinoma/surgery
• Contrast Media
• Dextrans
• Feasibility Studies
• Female
• France
• Humans
• Magnetite Nanoparticles
• Magnetometry/instrumentation
• Middle Aged
• Prospective Studies
• Radiopharmaceuticals
• Sentinel Lymph Node Biopsy/methods

Sentinel lymph node (SLN) examination is a standard in breast cancer patients, with several methods employed along its 20 years history, the last one represented by one-step nucleic acid amplification (OSNA). The latter is a intra-operative molecular assay searching for CK19 mRNA as a surrogate of metastatic cells. Our 3 years experience with OSNA (1122 patients) showed results overlapping those recorded in the same institution with a morphological evaluation (930 patients) of SLN. In detail, the data of OSNA were almost identical to those observed with standard post-operative procedure in terms of patients with positive SLN (30%) and micrometastatic/macrometastatic involvement of SLN (respectively, 38–45 and 62–55%). By contrast, when OSNA was compared to the standard intraoperatory procedure, it was superior in terms of accuracy, prompting the use of this molecular assay as a very valid, and reproducible for intra-operative evaluation of SLN. Further possibilities prompting the use of OSNA range from adhesion to quality control programs, saving of medical time, ability to predict, during surgery, additional nodal metastasis, and molecular bio-banking.

• accuracy
• breast cancer
• frozen section
• molecular analysis
• morphological analysis
• permanent sections
• sentinel node examination

• histopathology
• lymph nodes
• medullary thyroid cancer
• one-step nucleic acid amplification

To apply an individualized oncological approach to gastric cancer patients, the accurate diagnosis of disease entities is required. Peritoneal metastasis is the most frequent mode of metastasis in gastric cancer, and the tumor-node-metastasis classification includes cytological detection of intraperitoneal cancer cells as part of the staging process, denoting metastatic disease. The accuracy of cytological diagnosis leaves room for improvement; therefore, highly sensitive molecular diagnostics, such as an enzyme immunoassay, reverse transcription polymerase chain reaction, and virus-guided imaging, have been developed to detect minute cancer cells in the peritoneal cavity. Molecular targeting therapy has also been spun off from basic research in the past decade. Although conventional cytology is still the mainstay, novel approaches could serve as practical complementary diagnostics to cytology in near future.

• Gastric cancer
• Peritoneal lavage
• Cytology
• Molecular diagnostic techniques
• Reverse transcriptase polymerase chain reaction
• Carcinoembryonic antigen

• Biomarkers, Tumor/genetics
• Flow Cytometry
• Genetic Predisposition to Disease
• Genetic Testing
• Humans
• Immunoassay
• Molecular Diagnostic Techniques
• Neoplasm Invasiveness
• Peritoneal Neoplasms/chemistry
• Peritoneal Neoplasms/diagnosis
• Peritoneal Neoplasms/genetics
• Peritoneal Neoplasms/secondary
• Peritoneal Neoplasms/therapy
• Phenotype
• Precision Medicine
• Predictive Value of Tests
• Prognosis
• Stomach Neoplasms/chemistry
• Stomach Neoplasms/diagnosis
• Stomach Neoplasms/genetics
• Stomach Neoplasms/pathology

• Adolescent
• Adult
• Aged
• Aged, 80 and over
• Biomarkers, Tumor/analysis
• Breast Neoplasms/pathology
• Breast Neoplasms/surgery
• Carrier Proteins/analysis
• Carrier Proteins/biosynthesis
• Female
• Glycoproteins/analysis
• Glycoproteins/biosynthesis
• Humans
• Intraoperative Period
• Keratin-19/analysis
• Keratin-19/biosynthesis
• Lymphatic Metastasis/diagnosis
• Membrane Transport Proteins
• Middle Aged
• Nucleic Acid Amplification Techniques/methods
• Sentinel Lymph Node Biopsy/methods
• Young Adult

• BREAST CANCER
• MOLECULAR PATHOLOGY
• SENTINEL NODE

• Breast Cancer
• Lymph Nodes
• Molecular Pathology
• Surgery

• breast neoplasms
• cancer, staging
• keratin-19
• nucleic acid amplification techniques
• sentinel lymph node biopsy

• stomach neoplasm
• lymphatic metastasis
• intraoperative period
• nucleic acid amplification techniques
• keratin-19

• Adenocarcinoma/genetics
• Adenocarcinoma/secondary
• Adenocarcinoma/surgery
• Adenocarcinoma, Mucinous/genetics
• Adenocarcinoma, Mucinous/secondary
• Adenocarcinoma, Mucinous/surgery
• Adult
• Aged
• Aged, 80 and over
• Biomarkers, Tumor/analysis
• Carcinoma, Papillary/genetics
• Carcinoma, Papillary/secondary
• Carcinoma, Papillary/surgery
• Carcinoma, Signet Ring Cell/genetics
• Carcinoma, Signet Ring Cell/secondary
• Carcinoma, Signet Ring Cell/surgery
• Female
• Follow-Up Studies
• Humans
• Lymphatic Metastasis
• Male
• Middle Aged
• Neoplasm Invasiveness
• Neoplasm Staging
• Nucleic Acid Amplification Techniques/methods
• Prognosis
• Sentinel Lymph Node Biopsy
• Stomach Neoplasms/genetics
• Stomach Neoplasms/pathology
• Stomach Neoplasms/surgery

• Breast cancer
• Cytokeratin 19 mRNA copy number
• OSNA
• One step nucleic acid
• Sentinel lymph node biopsy

• axilla
• breast cancer
• cytology
• hospital economics
• sentinel lymph node biopsy

• Breast
• Lymph nodes
• Nucleic acid amplification
• OSNA
• Sentinel

• CK19 mRNA
• OSNA
• PCR assay
• breast cancer
• intraoperative assay
• sentinel lymph node

• Adult
• Aged
• Aged, 80 and over
• Axilla
• Breast Neoplasms/metabolism
• Breast Neoplasms/pathology
• Breast Neoplasms/physiopathology
• Female
• Humans
• Immunohistochemistry
• In Situ Hybridization
• Keratin-19/genetics
• Lymph Nodes/metabolism
• Lymph Nodes/pathology
• Lymphatic Metastasis/genetics
• Lymphatic Metastasis/physiopathology
• Middle Aged
• Neoplasm Micrometastasis/genetics
• Neoplasm Micrometastasis/physiopathology
• Prospective Studies
• Sentinel Lymph Node Biopsy

The one-step nucleic acid amplification (OSNA) assay is a rapid procedure for the detection of lymph node (LN) metastases using molecular biological techniques. The aim of this study was to assess the reliability of the whole sentinel lymph node (SLN) analysis by the OSNA assay as a predictor of non-SLN metastases.

Consecutive 742 patients with breast cancer were enroled in the study. The association of non-SLN or ⩾4 LN metastases with clinicopathological variables was investigated using multivariate logistic analysis.

In total, 130 patients with a positive SLN who underwent complete axillary LN dissection were investigated. The frequency of non-SLN metastases in patients who were OSNA+ and ++ was 19.3% and 53.4%, respectively, and that in patients with ⩾4 LN metastases who were OSNA+ and ++ was 7.0% and 27.4%, respectively. The cytokeratin 19 (CK19) mRNA copy number (⩾5.0 × 10³; OSNA++) in the SLN was the most significant predictors of non-SLN metastases (P=0.003). The CK19 mRNA copy number (⩾1.0 × 10⁵) in the SLN was the only independent predictor of ⩾4 LN metastases (P=0.014).

Whole SLN analysis using the OSNA assay could become a valuable method for predicting non-SLN and ⩾4 LN metastases.

Analysis of sentinel lymph node (SLN) by means of One-Step Nucleic Acid Amplification (OSNA) is being used increasingly as a very sensitive and quick method for intraoperative axillary staging in patients with breast cancer. This molecular diagnostic assay detects the expression level of cytokeratin 19 (CK19), a luminal epithelial cell marker broadly expressed in most breast carcinomas and not normally found in lymph nodes. Almost all breast cancers express this cytoskeleton protein, but some breast tumors have been found to lose the expression of CK19. CK19 immunostaining in core biopsies has been recommended in selecting patients eligible for OSNA analysis because SLNs with metastatic involvement by CK19-negative breast cancers may result in a false negative result by OSNA. However, the real frequency of CK19-negative breast cancer has to be elucidated. In this study, we have assessed the frequency and molecular profile of CK19-negative breast carcinomas in three series of cases. The first is a prospective series of 197 breast carcinomas, 111 of which were subjected to SLN evaluation by OSNA. The second is a retrospective series of 41 triple-negative (TN) breast carcinomas, and the third is a retrospective series of 68 breast cancer patients (matched core biopsies and metastatic lymph nodes) that had been evaluated by conventional procedures before the OSNA methodology was adopted in our institution. Our results not only demonstrate that lack of expression of CK19 is infrequent in breast cancers but also that performing CK19 immunohistochemical staining is important on diagnostic core biopsies in taking the decision of using OSNA methodology in the evaluation of sentinel nodes in breast cancer patients.