Early detection of human cytomegalovirus can prevent se-rious health consequences for immunocompromised pa-tients. Therefore, it is important to employ methods of sen-sitive and accurate detection. In this paper, we comparedthe sensitivity and specificity of detection of three methods:1) COBAS Amplicor ™ CMV MONITOR Test (CACM), com-mercial and validated method; it uses polymerase chainreaction (PCR) to detect the early gene Pol, 2) real-timePCR, and 3) real-time reverse transcription coupled toPCR (RT-PCR); the 2 and 3 were homemade, which vali-dated the detection of immediate early gene Ie1. We per-formed a cross-sectional study with 42 positive HIV-1 hu-man plasma samples. Nucleic acids were extracted usingtwo methods: 1) a caotropic agent and 2) an affinity co-lumn. We obtained a 9.5, 26.2, and 33.3%of positive sam-ples with CACM, PCR, and RT-PCR, respectively. Usingthe techniques homemade, we detected a minimum of 26copies/mL. We used the kappa test to determine correla-tion between methods, obtaining values of 0.34 (PCR vs.CACM), 0.0079 (RT-PCR vs.CACM), and 0.65 (RT-PCR vs.PCR). The greater sensitivity and specificity were obtainedby real time PCR, as well as the largest positive and nega-tive predictive values. We concluded that the qualitativedetection of immediate early gene Ie1(synonymous of acti-ve infection) of HCMV in positive HIV-1 human plasmasamples by real time PCR could be used as an alternativemethod to CACM.