Droplet digital PCR assay for precise determination of FRS2 gene copy number in bladder cancer.

ABSTRACT:

Objective

To establish and validate a droplet digital PCR (ddPCR) assay for quantifying FRS2 gene copy number in formalin-fixed paraffin-embedded (FFPE) bladder cancer tissue samples, and to evaluate its analytical performance.

Methods

The ddPCR assay was developed using FRS2 as the target gene and RPP30 as the reference gene. Artificial plasmids, genomic DNA from urinary sediment of healthy individuals and cell lines were used as templates to assess the assay's precision, minimum reliable input DNA, and linearity. Fluorescence in situ hybridization (FISH) was employed to validate the accuracy of the ddPCR results.

Results

One-dimensional fluorescence amplitude plots showed clear separation between positive and negative droplets for both FRS2 and RPP30. Duplex detecti

SUBMITTER: Li J 

PROVIDER: S-EPMC12291395 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature