title: Mutational analysis of cytocentrifugation supernatant fluid from pancreatic solid mass lesions.
abstract: Diagnosis of fine-needle aspirations of pancreatic solid masses is complicated by many factors that keep its false-negative rate high. Our novel approach analyzes cell-free cytocentrifugation supernatant, currently a discarded portion of the specimen. Supernatant and cytology slides were collected from 25 patients: 11 cases with confirmed outcome [five positive (adenocarcinoma) and six negative (inflammatory states)], plus 14 without confirmed outcomes. Slides were microdissected, DNA was extracted from microdissections and corresponding supernatants, and all were analyzed for KRAS point mutation and loss of heterozygosity. Notably, higher levels of free DNA were found in supernatants than in corresponding microdissected cells. Supernatants contained sufficient DNA for mutational profiling even when samples contained few to no cells. Mutations were present in 5/5 malignancies and no mutations were present in inflammatory states. In conclusion, these findings support using supernatant for mutational genotyping when diagnostic confirmation is required for pancreatic solid masses.
authors.affiliation: RedPath Integrated Pathology, Inc., Pittsburgh, Pennsylvania, USA.
authors.forename: Sydney D
authors.initials: SD
authors.lastname: Finkelstein
authors.forename: Marluce
authors.initials: M
authors.lastname: Bibbo
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Kowalski
authors.forename: David E
authors.initials: DE
authors.lastname: Loren
authors.forename: Ali A
authors.initials: AA
authors.lastname: Siddiqui
authors.forename: Charalambos
authors.initials: C
authors.lastname: Solomides
authors.forename: Eric
authors.initials: E
authors.lastname: Ellsworth
mesh_terms: D001826:Body Fluids / Q000378:metabolism*; D002498:Centrifugation; D004252:DNA Mutational Analysis / Q000379:methods*; D004273:DNA, Neoplasm / Q000235:genetics; D006801:Humans; D042282:Microdissection; D010190:Pancreatic Neoplasms / Q000235:genetics*
chemical_list: D004273:DNA, Neoplasm
keywords: cytology supernatant; mutational analysis; pancreas