HistoMosaic Detecting KRAS G12V Mutation Across Colorectal Cancer Tissue Slices through in Situ PCR

We report on HistoMosaic, a novel technique for genetic analysis of formalin-fixed, paraffin-embedded tissue slices. It combines microfluidic compartmentalization, in situ allele-specific PCR, and fluorescence microscopy. The experimental proof of principle was achieved by in situ detection of KRAS...

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Bibliographic Details
Published inAnalytical chemistry (Washington) Vol. 88; no. 5; pp. 2792 - 2798
Main Authors Raub, Christopher B, Lee, Chen-Chung, Shibata, Darryl, Taylor, Clive, Kartalov, Emil
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 01.03.2016
Subjects
Analytical chemistry
Cancer
Colorectal Neoplasms - pathology
Construction
Economics
Fluorescence
Genes, ras
Genetics
Heterogeneity
Humans
Microscopy, Confocal
Mutation
Mutations
Patients
Polymerase chain reaction
Polymerase Chain Reaction - methods
Tissues
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Summary:We report on HistoMosaic, a novel technique for genetic analysis of formalin-fixed, paraffin-embedded tissue slices. It combines microfluidic compartmentalization, in situ allele-specific PCR, and fluorescence microscopy. The experimental proof of principle was achieved by in situ detection of KRAS G12V mutation in colorectal cancer tissues and is presented herein. HistoMosaic offers the ability to detect mutations over the entire tissue slide simultaneously, rapidly, economically, and without selection bias, while coregistering the genetic information with the preserved morphological information. Thus, HistoMosaic has wide applicability in basic science as a tool to map genetic heterogeneity. It is also a platform to build companion diagnostics for targeted therapies in oncology, to help ensure that the right drug is given to the right patient, thereby saving healthcare resources and improving patient outcomes.
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C.B.R.: Department of Biomedical Engineering, The Catholic University of America, 620 Michigan Avenue NE, Washington, DC, 20064, USA.
Author Contributions
The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.5b04460