Abstract
Human cytomegalovirus (hCMV) is a widespread herpesvirus proposed to exert oncomodulatory effects on several cancers by influencing cell proliferation, inflammation, and immune evasion. hCMV DNA and proteins have been detected in tumors, such as glioblastoma and colorectal cancer, suggesting a possible role in tumor biology. However, detection rates in clinical samples vary dramatically from 0 to 100%, reflecting both biological heterogeneity and methodological differences. Standard detection methods such as immunohistochemistry, qPCR, and sequencing provide good sensitivity, but each has limitations, including inconsistent antibody performance, costly instrumentation, and limited suitability for routine screening in point-of-care settings. Electrochemical (EC) and electrogenerated chemiluminescence (ECL) bioassays and biosensors offer attractive alternatives, providing rapid, simple, and low-sample-consumption assays that are well suited for point-of-care testing. These platforms combine high analytical sensitivity with minimal instrumentation requirements, short turnaround times, and compatibility with amplification strategies, making them promising tools for decentralized viral diagnostics. In this study, we optimized and compared EC and ECL bioassays coupled with various amplification strategies, including PCR, recombinase polymerase amplification (RPA), and loop-mediated isothermal amplification (LAMP), for hCMV detection in real samples. Amperometric EC readout combined with PCR or RPA showed high specificity, reliably distinguishing hCMV-infected from uninfected cells, and demonstrating its potential for use in tumor tissue analysis. LAMP offers excellent sensitivity but suffers from nonspecific amplification, whereas the ECL assay failed to detect viral DNA under our conditions. These results highlight the relative strengths and limitations of these platforms and indicate their suitability for future hCMV diagnostics.