Ultra-Rapid Molecular-Genetic Assays Feasible for Tumor Classification During Surgery
MONDAY, March 3, 2025 -- Intraoperative ultra-rapid droplet digital polymerase chain reaction (UR-ddPCR) can identify tumor genetic subtype and quantify tumor cell percentage with high accuracy, according to a study published online Feb. 25 in Med.
Zachary R. Murphy, Ph.D., from the New York University Grossman School of Medicine in New York City, and colleagues introduced UR-ddPCR to examine its feasibility for use during surgery and whether the workflow can reduce the time from tissue biopsy to molecular diagnosis.
The researchers developed a UR-ddPCR assay for the IDH1 R132H mutation that accurately measured the mutant DNA percentage down to 0.1 percent, which was concordant with standard ddPCR. In addition, a UR-ddPCR assay was developed for the BRAF V600E mutation. The clinical feasibility of UR-ddPCR was assessed by intraoperative use in 22 brain tumor cases, with multiple tissue samples per case (78 samples). The tissue samples were used for UR-Stimulated Raman Histology and UR-ddPCR. There was high concordance for tumor cell percentages measured by intraoperative UR-ddPCR with standard ddPCR performed on the sample lysates. Raman histologic images provided real-time measurement of total cellularity, with tumor cell densities ranging from >1,300 to <5 tumor cells/mm2 within the core and at tumor margins, respectively. The UR-ddPCR results were achieved in an average of 14 minutes and 39 seconds and 17 minutes and seven seconds for IDH1 R132H and BRAF V600E assays, respectively.
"Our study shows that ultra-rapid droplet digital PCR could be a fast and efficient tool for making a molecular diagnosis during surgery for brain cancer," co-senior author Gilad D. Evrony, M.D., Ph.D., also from the New York University Grossman School of Medicine, said in a statement.
Several authors disclosed ties to the biopharmaceutical and medical device industries; several authors are listed on a provision patent on the technology. Reagents and instruments were provided by Bio-Rad.
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