Evaluation of New Quantitative Assays for Diagnosis and Monitoring of Cytomegalovirus Disease in Human Immunodeficiency Virus-Positive Patients

Abstract

Cobas Amplicor CMV Monitor (CMM) and Quantiplex CMV bDNA 2.0 (CMV bDNA 2.0), two new standardized and quantitative assays for the detection of cytomegalovirus (CMV) DNA in plasma and peripheral blood leukocytes (PBLs), respectively, were compared to the CMV viremia assay, pp65 antigenemia assay, and the Amplicor CMV test (P-AMP). The CMV loads were measured in 384 samples from 58 human immunodeficiency virus (HIV) type 1-infected, CMV-seropositive subjects, including 13 with symptomatic CMV disease. The assays were highly concordant (agreement, 0.88 to 0.97) except when the CMV load was low. Quantitative results for plasma and PBLs were significantly correlated (Spearman ρ = 0.92). For PBLs, positive results were obtained 125 days before symptomatic CMV disease by CMV bDNA 2.0 and 124 days by pp65 antigenemia assay, whereas they were obtained 46 days before symptomatic CMV disease by CMM and P-AMP. At the time of CMV disease diagnosis, the sensitivity, specificity, and positive and negative predictive values of CMV bDNA 2.0 were 92.3, 97.8, 92.3, and 97.8%, respectively, whereas they were 92.3, 93.3, 80, and 97.8%, respectively, for the pp65 antigenemia assay; 84.6, 100, 100, and 95.7%, respectively, for CMM; and 76.9, 100, 100, and 93.8%, respectively, for P-AMP. Considering the entire follow-up, the sensitivity, specificity, and positive and negative predictive values of CMV bDNA 2.0 were 92.3, 73.3, 52.1, and 97.1%, respectively, whereas they were 100, 55.5, 39.4, and 100%, respectively, for the pp65 antigenemia assay; 92.3, 86.7, 66.7, and 97.5%, respectively, for CMM; and 84.6, 91.1, 73.3, and 95.3%, respectively, for P-AMP. Detection of CMV in plasma is technically easy and, despite its later positivity (i.e., later than in PBLs), can provide enough information sufficiently early so that HIV-infected patients can be effectively treated. In addition, these standardized quantitative assays accurately monitor the efficacy of anti-CMV treatment.

Diagnosis of cytomegalovirus infections by qualitative and quantitative PCR in HIV infected patients

“…In our study, CD 4 lymphocyte counts below 100 cells/mm 3 were observed in a small number of the HIV infected patients having CMV genome detected (12/82, 13%) and only one of these cases was confirmed as having CMV disease. Our data corroborate those reported by the other authors 4,14,15,17 .…”

“…This high prevalence of CMV infections in the adult population is probably also observed in the HIV infected population. However, despite a high incidence of CMV infections a great decrease of CMV disease it is probably due to the raised use of HAART 4,14,15,17 . In our study, CD 4 lymphocyte counts below 100 cells/mm 3 were observed in a small number of the HIV infected patients having CMV genome detected (12/82, 13%) and only one of these cases was confirmed as having CMV disease.…”

Differences between the quantitative antigenemia assay and the cobas amplicor monitor quantitative PCR assay for detecting CMV viraemia in bone marrow and solid organ transplant patients

“…CMV disease is a major cause of morbidity and mortality in transplant patients. The likelihood of CMV disease is related to the viral load [Cope et al, 1997] and has prompted the development of quantitative methods to assist in the diagnosis and monitoring of CMV viraemia [Barkholt et al, 1999;Ghisetti et al, 1999;Pellegrin et al, 1999;Caliendo et al, 2000;Sia et al, 2000]. Two methods that have been used to quantitate CMV in peripheral blood are quantitative antigenemia and quantitative PCR.…”

Improvement in Long-Term Renal Graft Survival due to CMV Prophylaxis with Oral Ganciclovir: Results of a Randomized Clinical Trial

“…Temporary prophylaxis with 5 mg/kg body weight intravenous ganciclovir per day was initially allowed for patients who could not take oral medication. All patients were regularly monitored for CMV viral load in plasma using Cobas ® Amplicor ® CMV‐Monitor (Roche Diagnostics GmbH, Mannheim, Germany), a quantitative CMV PCR test (19,20). Monitoring was performed once weekly at weeks 1–4, every 2 weeks at weeks 5–12, every month at weeks 13–24 and every 3 months at weeks 25–52 or additionally, as clinically indicated.…”