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Updated: Aug 5, 2026
Discovery of Metastatic Regulators using a Rapid and Quantitative Intravital Chick Chorioallantoic Membrane Model
Published on: February 3, 2021
Mitochondrial DNA digital PCR improves metastasis quantification in the chick chorioallantoic membrane (CAM) assay
Shari Aerts1, Niene Daelman2, Gwen Sys3
1Department of Morphology, Imaging, Orthopedics, Rehabilitation and Nutrition, Faculty of Veterinary Sciences, Ghent University, Ghent, Belgium; CRIG (Cancer Research Institute Ghent), ConSarGhent, Ghent University, Ghent, Belgium; VON (Veterinary Oncology Network) -CRIG (Cancer Research Institute Ghent), Ghent University, Ghent, Belgium; DIGPCR Core, Ghent University, Ghent, Belgium.
None:
The chick chorioallantoic membrane (CAM) assay is an in vivo metastasis model, yet traditional Alu-qPCR quantification suffers from non-specific background noise in avian tissue. To improve analytical specificity, we developed a species-specific digital PCR (dPCR) assay targeting the human mitochondrial locus MT-ND4. While Alu-qPCR generated false positives in chicken-only DNA, MT-ND4 dPCR provided high species specificity and robust relative quantification of human mitochondrial DNA across various embryonic organs. As a proof of concept in five embryos, organ-dependent differences in metastatic burden were detected (p < 0.001) with high technical precision. This MT-ND4 dPCR assay avoids false-positive overestimations, providing a specific tool for quantifying human metastatic burden in the CAM model.
