Preparation of Cell Blocks for Lung Cancer Diagnosis and Prediction: Protocol and Experience of a High-Volume Center

Kossakowski, Claudia; Morresi‐Hauf, Alicia; Schnabel, Philipp A.; Eberhardt, Ralf; Herth, Felix; Warth, Arne

doi:10.1159/000357068

Cited by 23 publications

(21 citation statements)

References 38 publications

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“…5 ± 1 TBNA per patient in our cohort. Therefore, we support a statement made by others that sufficient representative needle passes per sample per cell block are recommended to provide sufficient DNA [4,20]. …”

Section: Discussionsupporting

confidence: 87%

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Suitability of Small Bronchoscopic Tumour Specimens for Lung Cancer Genotyping

Dooms

Vliegen²,

Borght³

et al. 2014

Respiration

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Background: Biomarker-driven clinical trials in advanced non-small cell lung cancer (NSCLC) usually accept biopsy specimens only, as cytology specimens are supposed to be more challenging due to low neoplastic cell content and suboptimal DNA quantity. Objectives: We aimed to evaluate 2 aspects of bronchoscopic biopsy and cytology specimens: (1) the proportion of neoplastic cells and quantity of DNA extracted, and (2) the detection limit of the Scorpion amplification refractory mutation system on endoscopic samples obtained in daily clinical practice. Methods: We screened 679 patients with advanced-stage NSCLC for the presence of an activating EGFR mutation according to the guidelines of the European Society of Medical Oncology. Their diagnostic tumour tissue samples were characterized. A dilution experiment was performed to determine the minimal proportion of neoplastic cells for a reliable test result. Results: Surgical biopsies, bronchoscopic forceps biopsy samples and needle aspiration cytology specimens exhibited a median tumour cell proportion of 70 versus 30 versus 20% and a DNA quantity of 2,500 versus 1,610 versus 1,440 ng, respectively. The overall EGFR mutation rate was 11%, with no differences between different sample types. Dilution experiments showed that the detection limit depends on the type of mutation. A neoplastic cell content of at least 10 and 25% for exon 19 deletions and exon 21 L858R point mutation, respectively, was required for a true negative result. Conclusions: Bronchoscopic forceps biopsy and needle aspiration cytology specimens are suitable for accurate EGFR mutation analysis using single-gene quantitative real-time polymerase chain reaction. Technologies with a better analytical sensitivity are evolving and should consider these endoscopic tumour specimens.

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Section: Discussionsupporting

confidence: 87%

“…the analytical sensitivity of the test). Recent guidelines and recommendations have focused on standardized and evidence-based material processing, analysis and reporting of results [2,3,4]. …”

Section: Introductionmentioning

confidence: 99%

Suitability of Small Bronchoscopic Tumour Specimens for Lung Cancer Genotyping

Dooms

Vliegen²,

Borght³

et al. 2014

Respiration

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“…As a consequence, the diagnosis of NSCLC should include genotyping as well as subtype classification [1,14,19,20,21,22]. Although various bronchoscopic specimens are available for genotyping [23,24,25,26,27,28,29,30], formalin-fixed paraffin-embedded specimens have been most widely employed in clinical practice because of their easy use, long-time storage and low costs [14]. Although the feasibility and reliability of genotyping in formalin-fixed paraffin-embedded bronchoscopic specimens obtained with standard-size forceps are well-established [23], it remains unclear whether the specimens sampled with the small biopsy forceps are suitable for genotyping.…”

Section: Introductionmentioning

confidence: 99%

Feasibility and Accuracy of Molecular Testing in Specimens Obtained with Small Biopsy Forceps: Comparison with the Results of Surgical Specimens

Oki

Yatabe²,

Sasaki³

et al. 2015

Respiration

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Background: During bronchoscopy, small biopsy forceps are increasingly used for the diagnosis of peripheral pulmonary lesions. However, it is unclear whether the formalin-fixed paraffin-embedded specimens sampled with the small biopsy forceps are suitable for the determination of genotypes which become indispensable for the management decision regarding patients with non-small cell lung cancer. Objectives: The aim of this study was to evaluate the feasibility and accuracy of molecular testing in the specimens obtained with 1.5-mm small biopsy forceps. Methods: We examined specimens in 91 patients, who were enrolled in our previous 3 studies on the usefulness of thin bronchoscopes and given a diagnosis of non-small cell lung cancer by bronchoscopy with the 1.5-mm biopsy forceps, and then underwent surgical resection. An experienced pathologist examined paraffin-embedded specimens obtained by bronchoscopic biopsy or surgical resection in a blind fashion on epidermal growth factor receptor (EGFR) mutations, anaplastic lymphoma kinase (ALK) rearrangements and KRAS mutations. Results: Twenty-five (27%), 2 (2%) and 5 (5%) patients had an EGFR mutation, ALK rearrangement and KRAS mutation, respectively, based on the results in surgical specimens. EGFR, ALK and KRAS testing with bronchoscopic specimens was feasible in 82 (90%), 86 (95%) and 83 (91%) patients, respectively. If molecular testing was feasible, the accuracy of EGFR, ALK and KRAS testing with bronchoscopic specimens for the results with surgical specimens was 98, 100 and 98%, respectively. Conclusion: The results of molecular testing in the formalin-fixed paraffin-embedded specimens obtained with the small forceps, in which the genotype could be evaluated, correlated well with those in surgically resected specimens.

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“…The cellblock technique for IHC has been used for a variety of solid tumors, such as those of the lung, pancreas, and breast, as well as lymphoma [19,20,21]. Yung et al [22] reported on a ‘tissue coagulum clot' cell block technique.…”

Section: Discussionmentioning

confidence: 99%

Fine-Needle Aspiration with Immunohistochemistry Using a Modified Scrape Cell Block Technique for the Diagnosis of Thyroid and Parathyroid Nodules

Wuertz

Kresnik

Malle³

et al. 2016

Acta Cytologica

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Objective: To evaluate the diagnostic accuracy of a modified scrape cell block (SCB) technique in a large series of patients. The technique was especially developed and tested for fine-needle aspiration of thyroid and parathyroid nodules. Study Design: Eighty-two ultrasound-guided fine-needle aspiration specimens with the sonographic aspect of a thyroid (n = 33) or a possible parathyroid nodule (n = 49) were studied. Immunohistochemistry (IHC) was used on cell blocks containing plasma, thromboplastin, and selected 3-dimensional cell aggregates scraped off Papanicolaou-stained smears. Antibodies for chromogranin A, thyroglobulin, parathyroid hormone, calcitonin, and carcinoembryonic antibody (CEA) were used. In cases of reduced immunosensitivity or suspected metastases or rare primary tumors, additional IHC markers were employed. Results: Chromogranin A was expressed in all 28 parathyroid adenomas (PA), in 7 of 8 hyperplastic parathyroid glands, and in 13 of 14 medullary thyroid carcinomas (MTC). When combining positivity for chromogranin A and calcitonin/CEA, the specificity for the detection of MTC was 100%. Parathyroid hormone was expressed in 26 of 36 parathyroid nodules (72.2%). When combining follicular microarchitecture and expression of chromogranin A, the specificity for the detection of parathyroid tissue was 97%. Conclusion: With the modified SCB technique, accurate cytological diagnoses were obtained in 97.6% of 82 patients.

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Preparation of Cell Blocks for Lung Cancer Diagnosis and Prediction: Protocol and Experience of a High-Volume Center

Cited by 23 publications

References 38 publications

Suitability of Small Bronchoscopic Tumour Specimens for Lung Cancer Genotyping

Suitability of Small Bronchoscopic Tumour Specimens for Lung Cancer Genotyping

Feasibility and Accuracy of Molecular Testing in Specimens Obtained with Small Biopsy Forceps: Comparison with the Results of Surgical Specimens

Fine-Needle Aspiration with Immunohistochemistry Using a Modified Scrape Cell Block Technique for the Diagnosis of Thyroid and Parathyroid Nodules

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