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

Flexman, James; Kay, Ian; Fonte, Rebecca; Herrmann, Richard; Gabbay, Eli; Palladino, Silvano

doi:10.1002/jmv.1047

Cited by 25 publications

(17 citation statements)

References 19 publications

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“…Five studies found CMV DNA PCR monitoring (qualitative or quantitative) to be more sensitive than Ag assay, irrespective of performance with plasma, PBL, or WB (26,40,76,89,93). In only one report were leukocyte-based assays (Ag and PBL PCR assays) more sensitive than a plasma PCR, showing a higher number of patients with CMV reactivation, earlier positivity, and a more rapid decrease of viral load after the start of preemptive antiviral therapy (9).…”

Section: Cobas Amplicor CMV Dna (Monitor) Testmentioning

confidence: 99%

Prevention of Cytomegalovirus Disease in Recipients of Allogeneic Stem Cell Transplants

2003

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The main risk factors for cytomegalovirus (CMV) disease in recipients of allogeneic stem cell transplants (SCT) are recipient CMV seropositivity and acute graft-versus-host disease. Currently, two antiviral strategies, prophylactic or preemptive antiviral treatment, are used for prevention of CMV disease. Preemptive treatment is most favorable when short-term (14-day) treatment is applied. Several methods are available for monitoring of CMV reactivation. PCR-based CMV DNA detection assays are the most sensitive methods; however, the clinical benefit of this high sensitivity is unclear. Even more, there is lack of clarity whether PCR tests can better be performed with plasma, whole blood, or peripheral blood leukocyte samples. Recovery of a CMV-specific CD8+ cytotoxic-T-lymphocyte (CTL) response is necessary for preventing CMV reactivation and disease. Reconstitution of absolute CMV-specific CTL counts to values above 10 × 106 to 20 × 106 CTLs/liter is associated with protection from CMV disease. In the near future, preemptive therapy might be withheld in patients with CMV reactivation who are shown to have adequate CMV-specific cytotoxic T-cell levels. Antiviral therapy with (val)acyclovir has been studied only as prophylactic treatment for prevention of CMV infection. High-dose oral valacyclovir is more effective than acyclovir when used in addition to preemptive treatment of CMV reactivation with ganciclovir or foscarnet. Three antiviral drugs have been tested for preemptive therapy of CMV reactivation and/or treatment of CMV disease. Although intravenous ganciclovir is considered the drug of choice, foscarnet has similar efficacy and less toxicity, especially hematologic toxicity. Cidofovir has not been tested extensively, but so far the results are disappointing. Oral valganciclovir for preemptive treatment of SCT recipients is currently being studied. In addition to antiviral therapy, adoptive immunotherapy with CMV-specific cytotoxic T cells as prophylactic or preemptive therapy is a very elegant strategy; however, generation of these cells is expensive and time-consuming, and therefore the therapy is not available at every transplantation center. Magnetic selection of CMV-specific CD8+ T cells from peripheral blood by using HLA class I-peptide tetramers may be very promising, making this strategy more accessible

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Section: Cobas Amplicor CMV Dna (Monitor) Testmentioning

confidence: 99%

Prevention of Cytomegalovirus Disease in Recipients of Allogeneic Stem Cell Transplants

2003

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“…Although quantitative PCR methods are widely used in CMV diagnostics, the clinical correlation of the findings is not as clear as that with the pp65 assay. High viral loads, measured by quantitative DNA-PCR methods, have been shown to correlate with the appearance of symptomatic CMV infection in a similar way (3,4,7,12,17,21,24). As the tendency nowadays is towards replacement of the pp65 assay with PCR methods, the clinical use of these methods should be evaluated in parallel, not only in the diagnosis of CMV infection but also in the monitoring of individual transplant patients during an active infection.…”

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confidence: 92%

Monitoring of Viral Load by Quantitative Plasma PCR during Active Cytomegalovirus Infection of Individual Liver Transplant Patients

Piiparinen¹,

Höckerstedt

Lappalainen³

et al. 2002

J Clin Microbiol

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A quantitative PCR test, the Cobas Amplicor CMV Monitor, was used for the monitoring of viral load in the peripheral blood of 27 individual liver transplant patients and correlated with cytomegalovirus (CMV) pp65 antigenemia. Altogether, 243 specimens were analyzed. During the first 3 months, 20 patients showed PCR positivity which correlated with pp65 antigenemia. Of those, 13 patients developed symptomatic CMV infection 27 to 52 days after transplantation, with a significantly higher peak viral load in PCR and in pp65 assay compared with the seven asymptomatic infections (median 10,200 versus 2,240 copies/ml, P < 0.05, and median 100 versus 30 pp65-positive cells/50,000 leukocytes, P < 0.01). Five were primary infections of D؉/R؊ cases (donor CMV seropositive and recipient seronegative) and demonstrated, except in one case, a high peak viral load (>10,000 copies/ml; range, 10,200 to 21,600 copies, and >50 positive cells, range, 50 to 800 cells). The peak viral loads of the six D؉/R؉ patients with symptomatic infection varied widely (range, 2,290 to 126,000 copies and 50 to 300 positive cells). Two D؊/R؉ patients developed symptomatic infection with a lower viral load (range, 1,120 to 6,510 copies and 25 to 100 positive cells). All symptomatic infections were successfully treated with ganciclovir. The asymptomatic infections all in D؉/R؉ patients with low copy numbers (<5,500 copies) were monitored until CMV disappeared. One of the seven PCR-negative patients had one sample with low antigenemia, but the subsequent specimens were all negative. The time-related correlation of the two methods was also good. In summary, quantitative PCR could equally well be used as the CMV pp65 assay for the monitoring of viral load in individual transplant patients.Cytomegalovirus (CMV) infection is a common complication after liver transplantation. A variety of clinical manifestations of CMV, such as fever, leukopenia, thrombocytopenia, colitis, pneumonia, and hepatitis, have been described (16). Immunosuppressed organ transplant patients are usually frequently monitored for CMV and treated with antiviral agents. The antiviral treatment is based on the clinical symptoms and/or rapid laboratory diagnosis. Also, preemptive therapy, which is commonly used to prevent the development of CMV disease after transplantation, is based on monitoring the viral load of the patients (5,6,15,20).For more than 10 years, the CMV-pp65 antigenemia assay has been the most commonly used method for monitoring the appearance of CMV infection in transplant patients (2,8,22,25). The antigenemia test is quantitative and can also be used to assess viral load and to monitor the response to antiviral treatment. Although there have been attempts to standardize the assay (8, 23), the great variety of in-house and commercial modifications of the method make it difficult to compare the results and run clinical trials based on this technique. Quantitative PCR techniques, which are easy to standardize, are less laborious for laboratory personnel, and can be autom...

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“…In fact, discrepant results have been published on this subject. (1,4,6,9,10,19,21,27,31,35,38,39,40). Recent data on the performance of several highly sensitive real-time PCRs, both laboratory-developed and commercial assays (5,20,22), however, demonstrate that both plasma and whole blood are equally suitable for monitoring active CMV infection in Allo-SCT recipients.…”

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confidence: 99%

Quantification of DNA in Plasma by an Automated Real-Time PCR Assay (Cytomegalovirus PCR Kit) for Surveillance of Active Cytomegalovirus Infection and Guidance of Preemptive Therapy for Allogeneic Hematopoietic Stem Cell Transplant Recipients

Gimeno

Solano

Latorre³

et al. 2008

J Clin Microbiol

108

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The performance of a plasma real-time PCR (cytomegalovirus [CMV] PCR kit; Abbott Diagnostics) was compared with that of the antigenemia assay for the surveillance of active CMV infection in 42 allogeneic hematopoietic stem cell transplantation (Allo-SCT) recipients. A total of 1,156 samples were analyzed by the two assays. Concordance between the two assays was 82.2%. Plasma DNA levels correlated with the number of pp65-positive cells, particularly prior to the initiation of preemptive therapy. Fifty-seven episodes of active CMV infection were detected in 37 patients: 18 were defined solely by the PCR assay and four were defined on the basis of the antigenemia assay. Either a cutoff of 288 CMV DNA copies/ml or a 2.42-log 10 increase of DNAemia levels between two consecutive PCR positive samples was an optimal value to discriminate between patients requiring preemptive therapy and those not requiring therapy on the basis of the antigenemia results. The real-time PCR assay allowed an earlier diagnosis of active CMV infection and was a more reliable marker of successful clearance of CMV from the blood. Analysis of the kinetics of DNAemia levels at a median of 7 days posttreatment allowed the prediction of the response to CMV therapy. Two patients developed CMV colitis. The PCR assay tested positive both before the onset of symptoms and during the disease period. The plasma real-time PCR from Abbott is more suitable than the antigenemia assay for monitoring active CMV infection in Allo-SCT recipients and may be used for guiding preemptive therapy in this clinical setting.Preemptive antiviral therapy based on the detection of cytomegalovirus (CMV) in blood has been shown to significantly reduce the incidence of CMV disease in the early posttransplant period following allogeneic hematopoietic stem cell transplantation (Allo-SCT) (2, 7). The pp65 antigenemia assay has been adopted by many transplant centers as the "gold standard" for monitoring active CMV infection and guiding preemptive therapy in this clinical setting. The antigenemia assay, however, requires rapid processing of specimens, is labor-intensive, cannot be automated, is subjective in reading, and is unfeasible during periods of severe neutropenia. In addition, it does not reflect the viral load in the blood compartment accurately (12) and sometimes displays negative results in the presence of CMV disease (2,31,32,36).Quantification of CMV DNA in blood by PCR is emerging as an alternative to the antigenemia assay and may soon become the standard for the surveillance of CMV infection in Allo-SCT recipients. Viral load quantification in blood allows early detection of active CMV infection, close monitoring of the response to antivirals, prediction of the risk of viremic relapse, emergence of resistant strains, and eventual development of CMV disease (8,31). In addition, it may be safely used for guiding preemptive therapy (11,16,23,24,32,37). Nevertheless, there is some controversy as to which is the optimal clinical specimen (plasma, whole blood, or leukoc...

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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

Cited by 25 publications

References 19 publications

Prevention of Cytomegalovirus Disease in Recipients of Allogeneic Stem Cell Transplants

Prevention of Cytomegalovirus Disease in Recipients of Allogeneic Stem Cell Transplants

Monitoring of Viral Load by Quantitative Plasma PCR during Active Cytomegalovirus Infection of Individual Liver Transplant Patients

Quantification of DNA in Plasma by an Automated Real-Time PCR Assay (Cytomegalovirus PCR Kit) for Surveillance of Active Cytomegalovirus Infection and Guidance of Preemptive Therapy for Allogeneic Hematopoietic Stem Cell Transplant Recipients

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