Vol. 9, Special Issue 10, Part M (2025)

Angiogenic biomarkers play important role in tumor detection. Soluble vascular endothelial growth factor-2 (sVEGFR-2) is one such marker and accurate detection of s-VEGFR-2 is crucial in tumor biology that help in early cancer diagnosis. Conventional PCR often fail to amplify low-copy transcripts which leads to false negatives. This study compares the performance of conventional PCR versus nested PCR to detect sVEGFR-2 in canine mammary tumors and is tallied with cDNA concentration on detection success. From histopatholoically confirmed thirty mammary tumor samples, RNA was extracted, reverse transcribed and conventional PCR as well as nested PCR was performed. Success of the amplification was compared in both the types of PCR and cDNA concentration was analysed relative to PCR positivity. A 546 bp amplicon was produced in 21 out of 30 (70%) tumor samples with conventional PCR, however, nested PCR produced 126 bp amplicon in 28 out of 30 (93.3%) tumor samples with seven additional cases that were negative with conventional PCR. Conventional PCR positives shown higher cDNA concentrations (mean 1024.3 ng/µl); whereas, nested PCR positives expanded detection to samples with lower template concentrations (mean 975.6 ng/µl). Nested PCR displayed superior sensitivity and specificity over conventional PCR. Nested PCR detected sVEGFR-2 transcripts from samples with cDNA concentrations.