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Mutational analysis of cytocentrifugation supernatant fluid from pancreatic solid mass lesions
Author(s) -
Finkelstein Sydney D.,
Bibbo Marluce,
Kowalski Thomas E.,
Loren David E.,
Siddiqui Ali A.,
Solomides Charalambos,
Ellsworth Eric
Publication year - 2014
Publication title -
diagnostic cytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.417
H-Index - 65
eISSN - 1097-0339
pISSN - 8755-1039
DOI - 10.1002/dc.23048
Subject(s) - genotyping , loss of heterozygosity , medicine , cytology , kras , adenocarcinoma , microbiology and biotechnology , dna , pathology , biology , genotype , genetics , gene , cancer , allele , colorectal cancer
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. Diagn. Cytopathol. 2014;42:719–725. © 2013 Wiley Periodicals, Inc.