Monitoring of cytomegalovirus reactivation after allogeneic stem cell transplantation: comparison of an antigenemia assay and quantitative real-time polymerase chain reaction

Abstract

Summary:Cytomegalovirus (CMV) antigenemia and quantitative real-time polymerase chain reaction (PCR) were compared for monitoring of CMV reactivation after allogeneic stem cell transplantation. The number of CMV antigen-positive cells by the antigenemia assay and the level of CMV DNA by real-time PCR correlated well. The sensitivity and specificity of the antigenemia assay was 55.4% and 95.5%, respectively, using real-time PCR as the reference standard. The probability of positive antigenemia at day 100 was 76.5%, with a median of first detection at day 37 in 51 patients, compared with a positive PCR of 84.3% and day 33, respectively. When HLA-identical sibling donor transplant recipients and other donor transplant recipients were analyzed separately, there was no difference between the two tests. However, temporal patterns of first detection of CMV antigen-positive cells and CMV DNA differed between HLA-identical and alternative recipients; patients without CMV (29%) or with sporadic positive PCR results (14%) were more common in HLA-identical sibling transplants, whereas patients with simultaneous antigenemia and positive PCR occurred more in alternative transplants (48%). Two of 51 patients (4%) developed CMV colitis despite antigenemia-guided prophylaxis, but both were successfully treated with ganciclovir. Although PCR is more sensitive than antigenemia, both tests are useful in the early detection of CMV after allogeneic stem cell transplantation.

Influence of antithymocyte globulin dose on outcome in cytomegalovirus‐seropositive recipients of partially T cell‐depleted stem cell grafts from matched‐unrelated donors

“…Until April 2001, CMV monitoring was performed using the CMV pp65 antigenaemia assay, thereafter monitoring was based on a real-time TaqMan TM CMV DNA PCR. This method is known to be more sensitive (Machida et al, 2000;Griscelli et al, 2001;Yakushiji et al, 2002) compared with the antigenaemia assay, however, the incidence of CMV reactivations was still lower in the recent group compared with the historical group. In conclusion, CMVseropositive MUD recipients of partial TCD grafts treated since 1999 show survival rates comparable to CMVseronegative recipients.…”

Real-time PCR assays based on distinct genomic regions for cytomegalovirus reactivation following hematopoietic stem cell transplantation

“…Many reports propose more than 200 copies/ ml as a suitable cutoff point. 5,20 However, this cutoff point was defined according to the sensitivity limit of real-time PCR assays and there is the potential risk that patients could be overtreated, resulting in unnecessary drug side effects and unnecessary cost. Our analysis suggests that CMV viral loads of 500 copies/ml might prove a more suitable cutoff point for initiating antiviral therapy in patient groups at high risk of CMV disease using the ROC curve, as real-time PCR targeting US17-and UL65-PCR can detect CMV infection earlier than antigenemia and nested PCR assays.…”

“…This confirmed results published previously. 3,5 Moreover, we investigated the correlation between the CMV viral loads obtained from real-time PCR amplifying a distinct genomic region. Results for all combinations of genomic regions correlated well.…”

“…Many reports have described the advantages of real-time PCR assays in comparison to antigenemia or qualitative PCR in patients who have undergone SCT. [3][4][5] However, some unsolved problems remain, not least that the procedure remains unstandardized. Whereas antigenemia assays have been almost completely standardized using an anti-pp65 antibody, 6,7 real-time PCR assays have been carried out using a variety of procedures: the target region being amplified by real-time PCR assay differs in each report, some target the immediate early protein (IE), while others target glycoprotein B (gB) or the US17 region, 2,4,8 and primers and probes for the target region also differ.…”

Comparative analysis of a double primer PCR assay with plasma, leukocytes and antigenemia for diagnosis of active human cytomegalovirus infection in bone marrow transplant patients

“…Similar results have been already reported using real-time PCR. 9,17,18 In this study, the PCR assay was easily capable of detecting 500 HCMV genomes/ml of clinical samples without nested PCR or hybridization assays. The sensitivity of multiplex PCR assays for detection of HCMV DNA in clinical samples of renal transplant recipients has already been described.…”