title: The use of FTA cards for preserving unfixed cytological material for high-throughput molecular analysis.
abstract: BACKGROUND
  Novel high-throughput molecular technologies have made the collection and storage of cells and small tissue specimens a critical issue. The FTA card provides an alternative to cryopreservation for biobanking fresh unfixed cells. The current study compared the quality and integrity of the DNA obtained from 2 types of FTA cards (Classic and Elute) using 2 different extraction protocols ("Classic" and "Elute") and assessed the feasibility of performing multiplex mutational screening using fine-needle aspiration (FNA) biopsy samples.
  
  
  METHODS
  Residual material from 42 FNA biopsies was collected in the cards (21 Classic and 21 Elute cards). DNA was extracted using the Classic protocol for Classic cards and both protocols for Elute cards. Polymerase chain reaction for p53 (1.5 kilobase) and CARD11 (500 base pair) was performed to assess DNA integrity.
  
  
  RESULTS
  Successful p53 amplification was achieved in 95.2% of the samples from the Classic cards and in 80.9% of the samples from the Elute cards using the Classic protocol and 28.5% using the Elute protocol (P = .001). All samples (both cards) could be amplified for CARD11. There was no significant difference in the DNA concentration or 260/280 purity ratio when the 2 types of cards were compared. Five samples were also successfully analyzed by multiplex MassARRAY spectrometry, with a mutation in KRAS found in 1 case.
  
  
  CONCLUSIONS
  High molecular weight DNA was extracted from the cards in sufficient amounts and quality to perform high-throughput multiplex mutation assays. The results of the current study also suggest that FTA Classic cards preserve better DNA integrity for molecular applications compared with the FTA Elute cards.
authors.affiliation: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Mauro Ajaj
authors.initials: MA
authors.lastname: Saieg
authors.forename: William R
authors.initials: WR
authors.lastname: Geddie
authors.forename: Scott L
authors.initials: SL
authors.lastname: Boerner
authors.forename: Ni
authors.initials: N
authors.lastname: Liu
authors.forename: Ming
authors.initials: M
authors.lastname: Tsao
authors.forename: Tong
authors.initials: T
authors.lastname: Zhang
authors.forename: Suzanne
authors.initials: S
authors.lastname: Kamel-Reid
authors.forename: Gilda
authors.initials: G
authors.lastname: da Cunha Santos
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D044963:Biopsy, Fine-Needle; D016009:Chi-Square Distribution; D003581:Cytodiagnosis / Q000379:methods; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping / Q000379:methods*; D008297:Male; D008875:Middle Aged; D008967:Molecular Biology; D009367:Neoplasm Staging; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012494:Sampling Studies; D013048:Specimen Handling
chemical_list: D004273:DNA, Neoplasm