Introduction: CMV disease ranges from asymptomatic infection in immunocompetent people to severe and lifethreatening infections in neonates and immunocompromised patients. The present study was undertaken to study the seroprevalence of CMV and use of Real-time PCR in detection and quantification of CMV in immunocompromised patients. Material and Methods: A total of 57 samples were tested for CMV IgM and CMV IgG by ELISA technique. The detection and quantification of CMV DNA was done by real time PCR. Results: Out of 57samples, 6 (10.53%) tested positive for CMV IgM, 50 (87.72%) were positive for CMV IgG and 18 (31.58%) were positive for CMV DNA by PCR. Among 18 PCR Positive cases, 6 (33.334%) cases showed viral load of 1x10 3 IU/ml. 4 (22.22%) cases each showed viral load of 1x10 4 IU/ml, 1x10 5 IU/ml, 1x10 6 IU/ml respectively. There is no statistical association of age, gender and geographical distribution with CMV IgM, CMV IgG and CMV DNA positivity. (P>0.05). Conclusion: This study demonstrated that Real-time PCR is more reliable test than serological ELISA test in the diagnosis of cytomegalovirus as Real-time PCR detects and quantifies viral DNA which is useful in predicting the patient's risk for disease and monitoring the effect of antiviral therapy. Good hygiene and infection control practices are advised to prevent CMV transmission.