NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis
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The application of molecular diagnostic studies interrogating EGFR and KRAS mutations to stained cytologic smears of lung carcinoma.

Author(s): Betz BL, Roh MH, Weigelin HC, Placido JB, Schmidt LA, Farmen S, Arenberg DA, Kalemkerian GP, Knoepp SM

Publication: Am J Clin Pathol, 2011, Vol. 136, Page 564-71

PubMed ID: 21917678 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine if rapid Romanowsky-stained smears can be used as an alternative tissue source for mutation analysis through sequencing of Kirsten rat sarcoma viral oncogene homolog (KRAS) and epidermal growth factor receptor (EGFR).

Conclusion of Paper

All rapid Romanowsky-stained smears and formalin-fixed paraffin-embedded (FFPE) surgical blocks yielded sufficient DNA for mutation detection, but only 17 of the 26 FFPE blocks were sufficiently cellular for analysis. One FFPE cytological block contained only sparse clusters of adenocarcinoma cells resulting in a false negative result when mutation analysis was performed through sequencing KRAS and EGFR, but otherwise, mutation analysis between rapid Romanowsky-stained smears and FFPE blocks was concordant.

Studies

  1. Study Purpose

    The purpose of this study was to compare DNA mutation detection in rapid Romanowsky-stained cytological smears, surgical tissue scrapings, archived coverslipped cytological smears and FFPE tissue and cytological blocks obtained from lung tumors. DNA was extracted from rapid Romanowsky-stained smears using the pinpoint slide DNA isolation kit and from FFPE sections using the automated BioRobot EZI.

    Summary of Findings:

    All rapid Romanowsky-stained smears, including the 7 obtained by scraping adenocarcinoma tumors, the 5 fresh cytological smears, and the 21 coverslipped archived cytological smears, yielded sufficient DNA for mutation detection in KRAS and EGFR. Similarly, all 7 FFPE surgical blocks allowed for mutation detection, but only 17 of the 26 FFPE cytological blocks were sufficiently cellular for analysis. Concordant mutation analysis results were obtained for 23 of 24 paired specimens. Follow-up analysis of the discordant case indicated that the FFPE block contained only sparse clusters of adenocarcinoma cells resulting in a false negative.

    Biospecimens
    Preservative Types
    • Formalin
    • Other Preservative
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    DNA DNA sequencing
    Cell count/volume H-and-E microscopy
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    DNA sequencing Specific Targeted nucleic acid KRAS codon 12
    KRAS codon 13
    KRAS codon 61
    EGFR exon 19
    EGFR exon 21
    Biospecimen Preservation Type of fixation/preservation Air-dried
    Formalin (buffered)
    DNA sequencing Specific Type of tissue stain None
    Rapid Romanowsky
    Biospecimen Acquisition Method of cell acquisition Cytological smear
    Biospecimen Acquisition Method of tissue acquisition Surgical resection
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