title: Efficiency of EGFR mutation analysis for small microdissected cytological specimens using multitech DNA extraction solution.
abstract: BACKGROUND
  The microdissection method has greatly facilitated the isolation of pure cell populations for accurate analysis of mutations. However, the absence of coverslips in these preparations leads to poor resolution of cellular morphological features. In the current study, the authors developed the MultiTech DNA extraction solution to improve the visualization of cell morphology for microdissection and tested it for the preservation of morphological properties of cells, quality of DNA, and ability to detect mutations.
  
  
  METHODS
  A total of 121 cytological samples, including fine-needle aspirates, sputum, pleural fluid, and bronchial washings, were selected from hospital archives. DNA extracted from microdissected cells was evaluated by epidermal growth factor receptor (EGFR) mutation analysis using pyrosequencing, Sanger sequencing, and peptide nucleic acid (PNA)-mediated real-time polymerase chain reaction clamping. Morphological features of cells as well as DNA quality and quantity were analyzed in several cytological samples to assess the performance of the MultiTech DNA extraction solution. The results were compared with previous EGFR mutation tests.
  
  
  RESULTS
  The MultiTech DNA extraction solution improved the morphology of archived stained cells before microdissection and provided a higher DNA yield than the commercial QIAamp DNA Mini Kit in samples containing a minimal number of cells (25-50 cells). The authors were able to detect identical EGFR mutations by using different analysis platforms and consistently identified these mutations in samples comprising as few as 25 microdissected cells.
  
  
  CONCLUSIONS
  The MultiTech DNA extraction solution is a reliable medium that improves the resolution of cell morphology during microdissection. It was particularly useful in EGFR mutations of samples containing a small number of cells.
authors.affiliation: Department of Animal Biotechnology, College of Animal Bioscience and Technology, Konkuk University, Seoul, Korea.
authors.forename: Seo Young
authors.identifier: http://orcid.org/0000-0002-2466-0068
authors.initials: SY
authors.lastname: Oh
authors.affiliation: Department of Animal Biotechnology, College of Animal Bioscience and Technology, Konkuk University, Seoul, Korea.
authors.forename: Hoon Taek
authors.initials: HT
authors.lastname: Lee
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000473:pathology; D044963:Biopsy, Fine-Needle; D016172:DNA Fingerprinting / Q000379:methods; D004252:DNA Mutational Analysis / Q000379:methods*; D004273:DNA, Neoplasm / Q000235:genetics; D016208:Databases, Factual; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D008297:Male; D042282:Microdissection / Q000379:methods; D008967:Molecular Biology; D009154:Mutation / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012494:Sampling Studies; D012680:Sensitivity and Specificity
chemical_list: D004273:DNA, Neoplasm; D066246:ErbB Receptors
keywords: DNA quality; MultiTech DNA extraction solution; cell morphological quality; epidermal growth factor receptor (EGFR) mutation analysis; microdissection