Detection of <i>EGFR</i> and <i>KRAS</i> gene mutations using suspension liquid-based cytology specimens in metastatic lung adenocarcinoma

Zhao, Huan; Qiu, Tian; H, Guo; Ying, Jianming; Li, Junling; Zhang, Zhihui

doi:10.18632/oncotarget.22530

Cited by 13 publications

(10 citation statements)

References 26 publications

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“…However, even when the sampling is not cytological but histological (e.g., endobronchial forceps biopsy, core needle biopsy) the agile cytopathologist can employ quick cytological techniques for rapid on-site evaluation and specimen triage [ 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 ]. Small biopsies can be used to prepare the whole array of cytological specimens in several ways.…”

Section: Discussionmentioning

confidence: 99%

Testing EGFR with Idylla on Cytological Specimens of Lung Cancer: A Review

Caputo

D'Ardia

Sabbatino

et al. 2021

IJMS

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The current standard of care for advanced non-small-cell lung cancer is based on detecting actionable mutations that can benefit from targeted therapy. Comprehensive genetic tests can have long turn-around times, and because EGFR mutations are the most prevalent actionable mutation, a quick detection would enable a prompt initiation of targeted therapy. Furthermore, the scarcity of diagnostic material means that sometimes only cytologic material is available. The Idylla™ EGFR assay is a real-time PCR–based method able to detect 51 EGFR mutations in 2.5 h. Idylla is validated for use only on FFPE sections, but some researchers described their experiences with cytological material. We reviewed the relevant literature, finding four articles describing 471 cases and many types of cytological input material: smears, cell-block sections, suspensions, and extracted DNA. The sensitivity, specificity, and limit of detection appear comparable to those obtained with histological input material, with one exception: the usage of scraped stained smears as input may reduce the accuracy of the test. In conclusion, usage of cytological material as input to the Idylla EGFR test is possible. A workflow where common mutations are tested first and fast, leaving rarer mutations for subsequent comprehensive profiling, seems the most effective approach.

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Section: Discussionmentioning

confidence: 99%

Testing EGFR with Idylla on Cytological Specimens of Lung Cancer: A Review

Caputo

D'Ardia

Sabbatino

et al. 2021

IJMS

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“…All the materials in the current study were fixed using CytoRich Red, and these fixed materials appeared to be suitable for molecular detection studies. The ThinPrep method (Hologic, Bedford, Massachusetts) has also recently been used to determine the suitability of LBC using other fixative techniques for molecular testing in our institution. Furthermore, we should examine the optimal preparation conditions for diagnosis and molecular testing in a clinical setting, and further studies are needed to compare the results of detection using bronchoscopic LBC materials with the results of the current study.…”

Section: Discussionmentioning

confidence: 99%

“…28 All the materials in the current study were fixed using CytoRich Red, and these fixed materials appeared to be suitable for molecular detection studies. The ThinPrep method (Hologic, Bedford, Massachusetts) 29 has also recently been used to determine the suitability of LBC using other fixative techniques for ships between radiological findings on CT and PET and EGFR or KRAS gene mutational status, but found no correlations. 30,31 In contrast, Chiu et al 32 reported that solid ADCs had a significantly higher SUVmax than other subtypes (papillary, acinar, and lepidic) and that tumors with a solid growth pattern showed a significantly higher SUVmax than those without a solid growth pattern.…”

Section: Discussionmentioning

confidence: 99%

Genotyping and cytomorphological subtyping of lung adenocarcinoma based on liquid‐based cytology

Tanaka

Sakamoto

Suzuki

et al. 2019

Diagnostic Cytopathology

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Background Liquid‐based cytology (LBC) samples allow immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and molecular testing of nucleic acids to be performed in the remaining fixed cells. The current study aimed to examine the relationship between gene mutational status and cytomorphological features in primary lung adenocarcinoma (ADC) using LBC materials. Methods Forty consecutive patients with primary lung ADC underwent surgical resection in our hospital. Cytological material was obtained by scraping the cut‐surface of the lesion, and samples were fixed and stored as LBC materials using CytoRich Red. Epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations, anaplastic lymphoma kinase (ALK), and c‐ros oncogene 1 (ROS1) gene rearrangements were detected, and cytomorphological studies were performed. Results Twenty cases (50%) were positive for EGFR mutation and four (10%) were positive for KRAS mutation. ALK gene rearrangement was identified in one case (2.5%) by IHC and FISH, and ROS1 gene rearrangement was identified in one case (2.5%) by IHC and real‐time polymerase chain reaction. The KRAS‐positive group included higher proportions of cases with an inflammatory background (100%), predominantly papillary architecture (75%), and papillary‐type ADC pattern (75%) compared with the EGFR‐positive group and the other group, which included ALK and ROS1 gene rearrangements. Conclusions LBC material is suitable for use in molecular testing. Differences in major gene aberrations detected by this method might predict specific cytomorphological features.

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“…Liquid-based cytology (LBC) is a useful method for the cytological diagnosis-based screening of lung cancer using sputum and bronchoalveolar lavage, because of its high precision and the availability of automatic standard systems that combine morphological assessment and molecular analyses [4][5][6][7][8][9]. The remaining fixed cells can be used for molecular analysis by extracting nucleic acids such as DNA or RNA for advanced studies.…”

Section: Introductionmentioning

confidence: 99%

Molecular Analysis of Liquid-Based Cytological Specimen Using Virtually Positive Sputum with Adenocarcinoma Cells

et al. 2020

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Liquid-based cytology (LBC) analysis of sputum is a useful diagnostic and prognostic tool for detecting lung cancer. DNA and RNA derived from lung cancer cells can be used for this diagnosis. However, the quality of cytological material is not always adequate for molecular analysis due to the effect of formalin in the commercially available fixation kits. In this study, we examined DNA and RNA extraction methods for LBC analysis with formalin fixation, using lung carcinoma cell lines and sputum. The human non-small cell lung cancer cell lines were fixed with LBC fixation reagents, such as CytoRich red preservative. Quantification of thyroid transcription factor-1 (TTF-1) and actin mRNA, epidermal growth factor receptor (EGFR) DNA in HCC827, H1975, and H1299 cells, and mutation analysis of EGFR in HCC827 and H1975 cells were performed by quantitative PCR (qPCR) and fluorescence resonance energy transfer (FRET)-based preferential homoduplex formation assay (F-PHFA) method, respectively. mRNA and DNA extracted from cell lines using RNA and/or DNA extraction kits for formalin-fixed paraffin-embedded (FFPE) fixed with various LBC solutions were efficiently detected by qPCR. The detection limit of EGFR mutations was at a rate of 5% mutated positive cells in LBC. The detection limit of the EGFR exon 19 deletion in HCC827 was detected in more than 1.5% of the positive cells in sputum. In contrast, the detection limit of the T790M/L858R mutation in H1975 was detected in more than 13% of the positive cells. We also detected EGFR mutations using next generation sequencing (NGS). The detection limit of NGS for EGFR mutation was lower than that of the F-PHFA method. Furthermore, more than 0.1% of positive cells could be cytomorphologically detected. Our results demonstrate that LBC systems are powerful tools for cytopathological and genetic analyses. However, careful attention should be paid to the incidence of false negative results in the genetic analysis of EGFR mutations detected by LBC.

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Detection of EGFR and KRAS gene mutations using suspension liquid-based cytology specimens in metastatic lung adenocarcinoma

Cited by 13 publications

References 26 publications

Testing EGFR with Idylla on Cytological Specimens of Lung Cancer: A Review

Testing EGFR with Idylla on Cytological Specimens of Lung Cancer: A Review

Genotyping and cytomorphological subtyping of lung adenocarcinoma based on liquid‐based cytology

Molecular Analysis of Liquid-Based Cytological Specimen Using Virtually Positive Sputum with Adenocarcinoma Cells

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