Improvement of cellularity on cell block preparations using the so‐called tissue coagulum clot method during endobronchial ultrasound‐guided transbronchial fine‐needle aspiration

Yung, Rex; Otell, Susan; Illei, Peter B.; Clark, Douglas P.; Feller‐Kopman, David; Yarmus, Lonny; Askin, Frederic B.; Gabrielson, Edward; Li, Qing Kay

doi:10.1002/cncy.20199

Cited by 90 publications

(106 citation statements)

References 33 publications

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“…For example, treatment of patients with non-small cell carcinoma of the lung depends on the histological type (adenocarcinoma versus squamous cell carcinoma) and the mutational status of the epidermal growth factor receptor (EGFR) and KRAS genes. 1 Thus, adequate tissue sampling is essential not only for pathological diagnosis but also for molecular testing that is required to guide therapeutic decisions. [1][2][3] In clinical practice, minimally invasive fine needle aspiration (FNA) is a helpful and convenient method for establishing the diagnosis of solid tumors.…”

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confidence: 99%

“…1 Thus, adequate tissue sampling is essential not only for pathological diagnosis but also for molecular testing that is required to guide therapeutic decisions. [1][2][3] In clinical practice, minimally invasive fine needle aspiration (FNA) is a helpful and convenient method for establishing the diagnosis of solid tumors. FNA procedures are included in the recommended guidelines for the diagnosis of thyroid carcinomas, 4,5 lung carcinomas 6,7 and sarcomas, such as Ewing Sarcoma/primitive neuroectodermal tumor.…”

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confidence: 99%

“…If the tumor is locally advanced or metastatic, surgical resection is not performed. 1,6 In some patients, tumor size, location or co-morbid conditions may preclude concurrent core needle or excisional biopsy. 1 Therefore, efficient strategies and novel technologies are needed to apply molecular testing to FNA samples.…”

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confidence: 99%

“…1,6 In some patients, tumor size, location or co-morbid conditions may preclude concurrent core needle or excisional biopsy. 1 Therefore, efficient strategies and novel technologies are needed to apply molecular testing to FNA samples. Studies have attempted to improve the yield of DNA from cytological samples by using Fast Technology for Analysis cards for high-quality DNA preservation.…”

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confidence: 99%

“…9 Preparation of cell blocks using the tissue coagulum clot method has also increased the efficacy of tissue extraction from endobronchial and transbronchial FNA samples. 1 Additionally, use of COLD-PCR/direct sequencing for EGFR and KRAS detection in lung aspirates and locked nucleic acid-based PCR/sequencing assay in thyroid aspirates has improved the sensitivity of mutation detection. 10,12 Massively parallel or next-generation sequencing (NGS) technology is increasingly being used for mutational analysis of tumors for both clinical and research applications.…”

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Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

et al. 2014

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Increasing use of fine needle aspiration for oncological diagnosis, while minimally invasive, poses a challenge for molecular testing by traditional sequencing platforms due to high sample requirements. The advent of affordable benchtop next-generation sequencing platforms such as the semiconductor-based Ion Personal Genome Machine (PGM) Sequencer has facilitated multi-gene mutational profiling using only nanograms of DNA. We describe successful next-generation sequencing-based testing of fine needle aspiration cytological specimens in a clinical laboratory setting. We selected 61 tumor specimens, obtained by fine needle aspiration, with known mutational status for clinically relevant genes; of these, 31 specimens yielded sufficient DNA for next-generation sequencing testing. Ten nanograms of DNA from each sample was tested for mutations in the hotspot regions of 46 cancer-related genes using a 318-chip on Ion PGM Sequencer. All tested samples underwent successful targeted sequencing of 46 genes. We showed 100% concordance of results between next-generation sequencing and conventional test platforms for all previously known point mutations that included BRAF, EGFR, KRAS, MET, NRAS, PIK3CA, RET and TP53, deletions of EGFR and wild-type calls. Furthermore, next-generation sequencing detected variants in 19 of the 31 (61%) patient samples that were not detected by traditional platforms, thus increasing the utility of mutation analysis; these variants involved the APC, ATM, CDKN2A, CTNNB1, FGFR2, FLT3, KDR, KIT, KRAS, MLH1, NRAS, PIK3CA, SMAD4, STK11 and TP53 genes. The results of this study show that next-generation sequencing-based mutational profiling can be performed on fine needle aspiration cytological smears and cell blocks. Next-generation sequencing can be performed with only nanograms of DNA and has better sensitivity than traditional sequencing platforms. Use of next-generation sequencing also enhances the power of fine needle aspiration by providing gene mutation results that can direct personalized cancer therapy.

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Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

et al. 2014

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Cell Block Preparation Techniques and Applications in Veterinary Medicine

Walsh¹,

Patel²

2020

Veterinary Cytology

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Identification of EGFR mutation, KRAS mutation, and ALK gene rearrangement in cytological specimens of primary and metastatic lung adenocarcinoma

Cai

Wong

Chhieng

et al. 2013

Cancer Cytopathology

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BACKGROUND:The identification of molecular alterations has an important therapeutic implication in patients with lung adenocarcinomas. In the current study, the authors evaluated their experience with the identification of epidermal growth factor receptor (EGFR), Kirsten rat sarcoma viral oncogene homolog (KRAS) mutation, and anaplastic lymphoma kinase (ALK) gene rearrangement using cytological specimens of primary and metastatic lung adenocarcinoma. METHODS: A total of 54 cases of lung adenocarcinomas (11 primary and 43 metastatic tumors) in which molecular tests were performed were retrieved. Molecular tests were performed on the cell block material of 19 effusions and 35 fine-needle aspirates. EGFR mutation was evaluated by polymerase chain reaction sequencing analysis of exons 18, 19, 20, and 21. KRAS mutation was tested using polymerase chain reaction-single-strand conformational polymorphism analysis of codons 12 and 13. ALK gene rearrangement was evaluated by fluorescence in situ hybridization using an ALK break apart probe. RESULTS: Molecular tests were successful in 49 of 54 cases (91%). Evaluation of EGFR mutation, KRAS mutation, and ALK gene rearrangement were performed in 49 cases, 14 cases, and 22 cases, respectively. EGFR mutations were found in 14 of 49 cases (29%), including 5 primary and 9 metastatic tumors. Three metastatic/recurrent adenocarcinomas demonstrated an additional EGFR T790M mutation that was not identified in the original specimens. KRAS mutation was detected in 3 of 14 cases (21%) including 1 primary and 2 metastatic tumors. ALK gene rearrangement was evident in 3 of 22 cases (14%), all of which were metastatic tumors. CONCLUSIONS:The results of the current study have demonstrated the feasibility of using cytological specimens for EGFR mutation, KRAS mutation, and ALK gene rearrangement analysis. Repeating molecular testing in metastatic/ recurrent lung adenocarcinomas may uncover newly acquired molecular alterations. Cancer (Cancer Cytopathol) 2013;121:500-7.

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Improvement of cellularity on cell block preparations using the so‐called tissue coagulum clot method during endobronchial ultrasound‐guided transbronchial fine‐needle aspiration

Cited by 90 publications

References 33 publications

Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

Cell Block Preparation Techniques and Applications in Veterinary Medicine

Identification of EGFR mutation, KRAS mutation, and ALK gene rearrangement in cytological specimens of primary and metastatic lung adenocarcinoma

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