Cytomegalovirus (CMV) is a globally distributed herpesvirus that establishes lifelong latency and can cause severe disease in newborns, pregnant women and immunocompromised patients. Accurate laboratory diagnosis is essential to guide antiviral therapy, infection control, and epidemiological surveillance. Multiple diagnostic modalities are used, including nucleic acid amplification testing (NAAT) such as  polymerase chain reaction (PCR) and transcription-mediated amplification (TMA), clustered regularly interspaced short palindromic repeat (CRISPR-based assays), serology of Immunoglobulin  (IgM, IgG, IgG avidity), antigen detection (pp65), viral culture, dried blood spots (DBS), tissue biopsy, and functional CMV assays such as immunoassays (Flow cytometry, QuantiFERON-CMV  and ELISpot Assays) CMV smear are included. Molecular techniques, especially PCR-based methods, provide rapid, sensitive and specific detection of Cytomegalovirus DNA or RNA from various samples and remain the gold standard for congenital, transplant and immunodeficient patients. Serology and antigen-based testing provide complementary information, but have limitations in distinguishing active infection from latent infection. Recommended laboratory algorithms emphasize the integration of timely sampling, quantitative viral load monitoring, and patient immune status to optimize clinical decision making. Despite progress, there are still research gaps in universal newborn screening, prenatal prediction, standardization of viral load thresholds and analysis harmony. Future directions aim to improve molecular and immune-based diagnostics, standardize laboratory workflow, and improve patient care in high-risk populations.

CMV, congenital CMV, PCR, viral load, QuantiFERON-CMV, ELISpot, molecular diagnostics

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