Detection of ALK-Positive Non–Small-Cell Lung Cancers on Cytological Specimens: High Accuracy of Immunocytochemistry with the 5A4 Clone

Savic, Spasenija; Bode, Beata; Diebold, Joachim; Tosoni, Ivo; Barascud, Audrey; Baschiera, Betty; Grilli, Bruno; Herzog, Michelle; Obermann, Ellen C.; Bubendorf, Lukas

doi:10.1097/jto.0b013e3182936ca9

Cited by 89 publications

(101 citation statements)

References 27 publications

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“…Interestingly, the IHC 5A4 (Novocastra; Leica Biosystems, Buffalo Grove, IL, http://www. leicabiosystems.com) has been compared with FISH on a cytological specimen and showed sensitivity, specificity, PPV, and NPV of 93.3%, 96.0%, 93.3%, and 96%, respectively [28]; however, FISH served as the gold standard in this study. Our study supports the IHC-based screening approach for detecting EML4-ALK rearrangement in lung cancer.…”

Section: Discussionmentioning

confidence: 99%

Fluorescence In Situ Hybridization, Immunohistochemistry, and Next-Generation Sequencing for Detection of EML4-ALK Rearrangement in Lung Cancer

et al. 2015

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Background. The U.S. Food and Drug Administration-approved method for detecting EML4-ALK rearrangement is fluorescence in situ hybridization (FISH); however, data supporting the use of immunohistochemistry (IHC) for that purpose are accumulating. Previous studies that compared FISH and IHC considered FISH the gold standard, but none compared data with the results of next-generation sequencing (NGS) analysis. Materials and Methods. We studied FISH and IHC (D5F3 antibody) systematically for EML4-ALK rearrangement in 51 lung adenocarcinoma patients, followed by NGS in case of discordance. Results. Of 51 patients, 4 were positive with FISH (7.8%), and 8 were positive with IHC (15.7%). Three were positive with both. NGS confirmed that four of the five patients who were

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Section: Discussionmentioning

confidence: 99%

Fluorescence In Situ Hybridization, Immunohistochemistry, and Next-Generation Sequencing for Detection of EML4-ALK Rearrangement in Lung Cancer

et al. 2015

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“…Given the diversity of cytopathologic specimen types and the different methods of fixation and processing available, options for procuring DNA from cytologic specimens for molecular analysis vary; material can be successfully obtained from cell blocks, smears, cytospins, and liquid-based preparations. 7,9,11,12,14,[19][20][21][22][23][24][25] Most commonly, cell blocks are used for molecular testing on cytopathologic specimens. 21 The cell block closely approximates surgical pathology material; tissue fragments from cytopathologic specimens are embedded in paraffin and are sectioned akin to surgical specimens.…”

Section: Discussionmentioning

confidence: 99%

Molecular Testing of Non–Small Cell Lung Carcinoma Diagnosed by Endobronchial Ultrasound–Guided Transbronchial Fine-Needle Aspiration: The Cleveland Clinic Experience

Doxtader

Cheng

Zhang

2018

Archives of Pathology &Amp; Laboratory Medicine

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Context.— Given the increasing demand for molecular testing of non–small cell lung carcinoma specimens to guide therapeutic decision-making and the trend toward minimally invasive techniques for obtaining diagnostic tissue, cytopathology laboratories must devise strategies to maximize DNA yield for necessary molecular testing. Objective.— To describe our experience at Cleveland Clinic with epidermal growth factor receptor (EGFR) mutation testing by next-generation sequencing and anaplastic lymphoma kinase (ALK) gene rearrangement testing by fluorescence in situ hybridization of non–small cell lung carcinomas diagnosed by cytology, with an emphasis on specimens obtained by endobronchial ultrasound–guided transbronchial fine-needle aspiration. Data Sources.— Data sources include a review of the current literature, including published articles from our institution. Conclusions.— At our institution, liquid-based cytology specimens are the primary resource used for molecular testing of non–small cell lung carcinomas; in most instances, adequate DNA can be extracted from the residual cell pellet for next-generation sequencing, and ThinPrep slides can be used reliably for fluorescence in situ hybridization testing for ALK gene rearrangements. In occasional cases where the cell pellet material is not adequate for molecular testing, cell blocks and/or surgical pathology specimens are secondary options. The cytopathologist's role in specimen handling and triage is essential to ensure that molecular testing can be carried out successfully.

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“…Currently, FISH break-apart probe analysis is the gold standard for detection of ALK rearrangement and can be performed in cytology specimens. 17,44 However, it is an expensive, technically challenging test that is impractical to use on every specimen because the prevalence of ALK rearrangement is only approximately 5% among NSCLCs. 45,46 Although the first ALK antibody clones were unreliable, the subsequently developed clones D5F3 (Cell Signaling Technology) and 5A4 (Novocastra, Leica Biosystems, Buffalo Grove, Illinois) have a reported sensitivity of 100% and specificity of 99% in biopsy specimens, with excellent interobserver agreement and concordance with FISH results.…”

Section: Alk Antibodiesmentioning

confidence: 99%

“…45,46 Although the first ALK antibody clones were unreliable, the subsequently developed clones D5F3 (Cell Signaling Technology) and 5A4 (Novocastra, Leica Biosystems, Buffalo Grove, Illinois) have a reported sensitivity of 100% and specificity of 99% in biopsy specimens, with excellent interobserver agreement and concordance with FISH results. 6,7,17,26,47,48 Clone 5A4, which is as reliable as D5F3 in the detection of ALK rearrangement, is used more commonly in Europe. 17,49 The performance of ALK IPOX in cytology specimens (formalin-fixed cell blocks) thus far is similar to that in surgical specimens.…”

Section: Alk Antibodiesmentioning

confidence: 99%

“…Direct smears, cell monolayers, and cell block sections can all be used for IPOX and molecular tests including next-generation sequencing. 7,[13][14][15][16][17] IPOX TESTING Similar to core biopsies, low tumor content or DNA quality may be encountered in cytologic specimens despite safeguards in tissue acquisition, preparation, and use. 8 Immunoperoxidase ameliorates this limitation, as it detects signals in specimens with low cellularity.…”

Section: Cytology Specimensmentioning

confidence: 99%

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Lung Carcinoma Predictive Biomarker Testing by Immunoperoxidase Stains in Cytology and Small Biopsy Specimens: Advantages and Limitations

Zhou

Moreira²

2016

Archives of Pathology &Amp; Laboratory Medicine

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Context.-In the burgeoning era of molecular genomics, immunoperoxidase (IPOX) testing grows increasingly relevant as an efficient and effective molecular screening tool. Patients with lung carcinoma may especially benefit from the use of IPOX because most lung carcinomas are inoperable at diagnosis and only diagnosed by small tissue biopsy or fine-needle sampling. When such small specimens are at times inadequate for molecular testing, positive IPOX results still provide actionable information.Objective.-To describe the benefits and pitfalls of IPOX in the detection of biomarkers in lung carcinoma cytology specimens and small biopsies by summarizing the currently available commercial antibodies, preanalytic variables, and analytic considerations.Data Sources.-PubMed.Conclusions.-Commercial antibodies exist for IPOX detection of aberrant protein expression due to EGFR L858R mutation, EGFR E746_A750 deletion, ALK rearrangement, ROS1 rearrangement, and BRAF V600E mutation, as well as PD-L1 expression in tumor cells. Automated IPOX protocols for ALK and PD-L1 detection were recently approved by the Food and Drug Administration as companion diagnostics for targeted therapies, but consistent interpretive criteria remain to be elucidated, and such protocols do not yet exist for other biomarkers. The inclusion of cytology specimens in clinical trials would expand patients' access to testing and treatment, yet there is a scarcity of clinical trial data regarding the application of IPOX to cytology, which can be attributed to trial designers' lack of familiarity with the advantages and limitations of cytology. The content of this review may be used to inform clinical trial design and advance IPOX validation studies.

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Detection of ALK-Positive Non–Small-Cell Lung Cancers on Cytological Specimens: High Accuracy of Immunocytochemistry with the 5A4 Clone

Abstract: ALK ICC is highly accurate for detecting ALK-rearranged NSCLCs.

Cited by 89 publications

References 27 publications

Fluorescence In Situ Hybridization, Immunohistochemistry, and Next-Generation Sequencing for Detection of EML4-ALK Rearrangement in Lung Cancer

Fluorescence In Situ Hybridization, Immunohistochemistry, and Next-Generation Sequencing for Detection of EML4-ALK Rearrangement in Lung Cancer

Molecular Testing of Non–Small Cell Lung Carcinoma Diagnosed by Endobronchial Ultrasound–Guided Transbronchial Fine-Needle Aspiration: The Cleveland Clinic Experience

Lung Carcinoma Predictive Biomarker Testing by Immunoperoxidase Stains in Cytology and Small Biopsy Specimens: Advantages and Limitations

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