• A qPCR Multiplex Array assay for simultaneous detection of 21 viral sequences was developed. • The assay demonstrated high specificity, linearity (R 2 > 0.98), and reproducibility (CV 0.98), amplification efficiencies within the 90–110% range, and reproducibility (CV < 5%), confirming the reliability of this method for viral detection in clinical tissue samples. Viral sequences were detected in 30% (51/172) of cases for HPV, 24% (41/172) for MMTV, and 0.6% (1/172) for EBV. Sanger sequencing confirmed 100% sequence identity with GenBank reference sequences in all positive samples. A qPCR Multiplex Array assay targeting 21 viral sequences was developed and validated, providing a reliable tool for the detection of cancer-associated viral sequences in BC tissue. Detection of viral sequences confirms their presence in tumor tissue but does not establish causality with carcinogenesis. Future studies integrating transcriptomic, functional, and clinical analyses are needed to clarify the role of these viruses in BC, including associations with clinicopathological features and potential therapeutic implications.
Trujillo-Murillo et al. (Thu,) studied this question.