Multiplex Test Reveals Molecular Subtypes of Breast Cancer

Video protocol shows how to get robust results even from FFPE and other challenging samples

Sometimes there’s nothing better than watching someone else perform a protocol to understand how it’s done. That’s the concept behind JoVE, or the Journal of Visualized Experiments, which is kind of like YouTube for scientific techniques. In a new video and written publication, scientists from the University of Malta demonstrate a multiplex gene expression assay for the molecular classification of breast cancer tumors, particularly from formalin fixed paraffin-embedded (FFPE) samples.

“Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material” comes from Shawn Baldacchino, Christian Saliba, Jeanesse Scerri, Christian Scerri, and Godfrey Grech. The team’s 11-minute protocol video, which features the lead and senior authors, was produced through a grant from Luminex won by Godfrey. (The step-by-step protocol is also included in the written publication.)

For this project, scientists aimed to optimize an existing commercial kit for more robust use with archival FFPE or otherwise treated breast cancer tissue. They chose Invitrogen’s QuantiGene Plex Assay, which is based on our xMAP® Technology. “The gene expression assay described in this manuscript is a novel, quick, and multiplex method that can accurately classify breast cancer into the different molecular subtypes, omitting the subjectivity of interpretation inherent in imaging techniques,” the team reports. “This method has a wide range of possible applications including tumor classification with diagnostic potential and measurement of biomarkers in liquid biopsies, which would allow better patient management and disease monitoring.”

The video details important steps of the protocol, such as laser microdissection, lysis, and data analysis. Examples show the classification of tumors based on several well-known biomarkers into HER2-positive, triple negative, and other molecular subtypes of breast cancer.

Congratulations to Godfrey and his team on a great JoVE publication!

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Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material