Optimizing the <scp>DNA</scp> yield for molecular analysis from cytologic preparations

Roy‐Chowdhuri, Sinchita; Chow, Chi Wan; Kane, Mary K.; Yao, Hui; Wistuba, Ignacio I.; Krishnamurthy, Savitri; Stewart, John; Staerkel, Gregg

doi:10.1002/cncy.21664

Cited by 53 publications

(60 citation statements)

References 35 publications

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“…Dedicated passes for cell block only was left to the discretion of the thoracic physician performing the procedure. Although there is evidence for molecular testing on smears, the experience at our institution is that cell block provides better preservation of cells for testing (10,11). Currently our centre has switched to OncoFOCUS version 3 kit designed by AGENA for MALDI-TOF mass spectrometry which tests each tissue sample for EGFR, KRAS, NRAS, BRAF and KIT mutations.…”

Section: Methodsmentioning

confidence: 99%

Cell block samples from endobronchial ultrasound transbronchial needle aspiration provide sufficient material for ancillary testing in lung cancer—a quaternary referral centre experience

Hopkins¹,

Moffat²,

Parkinson³

et al. 2016

J. Thorac. Dis.

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Background: Rapid on site examination (ROSE) is encouraged at endobronchial ultrasound transbronchial needles aspiration (EBUS-TBNA) to improve diagnostic yield. Due to new therapeutic options in lung cancer, it is not sufficient to merely distinguish between non-small cell lung carcinoma (NSCLC) and small cell lung carcinoma (SCLC). Immunohistochemistry (IHC) distinction is now standard practice, as well as additional molecular testing where clinically indicated. We investigated the diagnostic yield of on-site smears vs. cell block and the provision of cellular material for ancillary testing at our centre.Methods: A retrospective audit of all EBUS-TBNA procedures performed until July 2012 was undertaken.Diagnostic yield on smears versus cell block was recorded. Cell blocks were reviewed by an experienced pathologist to determine diagnostic accuracy and whether IHC and molecular testing were possible. Results: In total, 234 procedures were recorded with 101 (43.2%) malignant cases, 107 (45.7%) benign cases and an initial 26/234 (11.1%) insufficient for diagnosis of which 11/234 (4.7%) were false negatives for malignancy after further follow up. The average number of passes was 4.5. For malignancies, smear diagnosis was possible in 95% (96/101) of cases and cell block diagnosis in 93.5% (87/93) of cases. There was sufficient material for IHC in 97.7% (85/87) of malignant cases. In 79.3% (69/87) of NSCLCs molecular testing for epidermal growth factor receptor (EGFR) mutation analysis was theoretically possible on samples obtained. Conclusions: Cell blocks are not inferior to smears for diagnostic accuracy and provide sufficient samples for histology. However, ROSE assists the physician on how best to manage samples for ancillary testing.

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Section: Methodsmentioning

confidence: 99%

Cell block samples from endobronchial ultrasound transbronchial needle aspiration provide sufficient material for ancillary testing in lung cancer—a quaternary referral centre experience

Hopkins¹,

Moffat²,

Parkinson³

et al. 2016

J. Thorac. Dis.

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“…19 Noteworthy, a significantly higher DNA yield was obtained with slides mounted with the low-hazard, organic, polymer-based mounting medium EcoMount (BioCare Medical LLC, Concord, California, USA) when compared with the xylene-based mounting medium Pertex (CellPath, Newtown, Powys, UK). 25 As technology is advancing at rapid pace, a range of novel techniques is emerging.…”

Section: Nucleic Acid Sample Quantity and Quality Assessmentmentioning

confidence: 99%

How to Prepare Cytological Samples for Molecular Testing

Bellevicine¹,

Malapelle²,

Vigliar³

et al. 2018

Molecular Applications in Cytology

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“…The type of slide used for cytology smears also influences DNA yield, which is significantly lower with fully frosted slides than with ordinary slides due to firm adherence of cells to the former [29] . We used only nonfrosted slides for FNA smears.…”

Section: Discussionmentioning

confidence: 99%

Use of Exfoliative Specimens and Fine-Needle Aspiration Smears for Mutation Testing in Lung Adenocarcinoma

Jain¹,

Ramachandrappa²,

Singh³

et al. 2017

Acta Cytologica

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Objective: Cytology specimens are considered to be a promising alternative for detecting driver mutations in lung cancer patients. We aimed to explore the suitability and utility of various cytology samples of non-small-cell lung cancer (NSCLC) patients for mutation testing. In addition to mutation detection, the importance of preanalytic factors was discussed. Design: A total of 116 cytology samples including 32 controls comprising pleural effusions, bronchial washings, and direct fine-needle aspiration (FNA) smears were included in the study for the detection of EGFR, KRAS, and Her-2/neu gene mutations. Tumor content was checked by microscopic evaluation. Tumor enrichment was done by scraping direct smears. DNA yield was assessed before selecting the method of mutation detection. Sanger sequencing and real-time PCR-based methods were used. Results: Overall, 20.23% EGFR mutations and 2.74% KRAS mutations were observed in this study. Nondriver genetic polymorphisms were observed in EGFR exon 20 in 37% cases. The coexistence of the EGFR mutation and EGFR polymorphism was seen in 7 cases. DNA yield was better in pleural effusions and bronchial washings. Real-time PCR was used in specimens of low DNA yield. Conclusions: Cytology samples are useful in ascertaining molecular diagnostic information for treatment purposes if they are optimized judiciously in their preanalytic phase.

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Optimizing the DNA yield for molecular analysis from cytologic preparations

Cited by 53 publications

References 35 publications

Cell block samples from endobronchial ultrasound transbronchial needle aspiration provide sufficient material for ancillary testing in lung cancer—a quaternary referral centre experience

Cell block samples from endobronchial ultrasound transbronchial needle aspiration provide sufficient material for ancillary testing in lung cancer—a quaternary referral centre experience

How to Prepare Cytological Samples for Molecular Testing

Use of Exfoliative Specimens and Fine-Needle Aspiration Smears for Mutation Testing in Lung Adenocarcinoma

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