Evaluation of the NucliSens CMV pp67 assay for detection and monitoring of human cytomegalovirus infection after allogeneic stem cell transplantation

Hebart, Holger; Rudolph, T; Loeffler, Juergen; Middeldorp, Jaap M.; Ljubicic, T; Jahn, Gerhard; Einsele, Hermann

doi:10.1038/sj.bmt.1703604

Cited by 25 publications

(6 citation statements)

References 23 publications

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“…It represents the centroid of the latter. ment (transplant recipients) [Pellegrin et al, 2000;Amorim et al, 2001;Kulkarni et al, 2001;Hebart et al, 2002]. However, the present report is the first concurrent quantitative real-time detection of CMV DNA by TaqMan method, pp65 antigenemia, and pp67 RNA assay.…”

Section: Discussionmentioning

confidence: 79%

Assessment of CMV load in solid organ transplant recipients by pp65 antigenemia and real‐time quantitative DNA PCR assay: Correlation with pp67 RNA detection

Mengoli

Cusinato

Biasolo

et al. 2004

Journal of Medical Virology

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After bone marrow (BM) or solid-organ (SO) transplantation viremic Cytomegalovirus (CMV) infection is observed frequently. Quantitative assay of CMV in blood helps the management of this clinical condition. In the present report, 83 samples from 39 solid organ recipients, three CMV assays were compared simultaneously for the first time: the Nuclisens CMV pp67 assay (nucleic acid sequence-based amplification, NASBA), an "in-house" quantitative real-time PCR assay (TaqMan) for CMV DNA, and pp65 antigenemia. The relation between CMV DNA and pp65 antigenemia, the quantitative assays, was evaluated on a larger group including 251 blood samples from 118 solid organ recipients. Real-time PCR provided the best results; > or =130 CMV DNA copies/2 x 10(5) peripheral blood leukocytes (PBLs) predicted > or =1 pp65 antigen positive (Ag+) cell/2 x 10(5) PBLs. By taking pp65 antigenemia as the "gold standard," the sensitivity of CMV DNA quantitation and of the pp67 RNA assay were 0.95 and 0.20, respectively, while the corresponding specificity values were 0.50 and 0.93. When real-time PCR was considered as the "gold standard," the sensitivity and specificity of the pp65 antigenemia were 0.65 and 0.91, respectively. Among the three tests examined, the sensitivity of the pp67 RNA assay was the lowest. On the other hand, the pp67 RNA assay was highly specific and effective in pinpointing high viremia patients. The present report, by providing predictive values for all three diagnostic profiles, DNA load, antigenemia, and pp67RNA, is a contribution for validation of real-time PCR as a new standard for quantitative assessment of CMV viremia in clinical settings.

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Section: Discussionmentioning

confidence: 79%

Assessment of CMV load in solid organ transplant recipients by pp65 antigenemia and real‐time quantitative DNA PCR assay: Correlation with pp67 RNA detection

Mengoli

Cusinato

Biasolo

et al. 2004

Journal of Medical Virology

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“…The qualitative determination of CMV pp67 mRNA by nucleic acid sequence-based amplification proved to be the least sensitive technique to assess CMV reactivation when compared to DNA-based assays and the Ag assay (28,39,76). Detection of immediate-early mRNA (29), the beta 2.7 transcript (1), or spliced late CMV genes (10) all were shown to be more useful; however, these results have not been validated by other groups.…”

Section: Mrna-based Monitoringmentioning

confidence: 98%

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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“…11,17 More recently, pp65-antigenemia assays and PCR methods have been established for HCMV monitoring of patients at risk. 14,18,19 Preemptive antiviral therapy based on pp65-antigenemia and HCMV-DNA PCR assays has been shown to significantly reduce both HCMV disease incidence and HCMV-associated mortality following allogeneic SCT in the early post transplant period, 11,12,[20][21][22][23] and has become a standard of care in many transplant centers. Preemptive ganciclovir therapy based on HCMV-pp65-antigenemia has resulted in an HCMV disease rate of approximately 14%.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the COBAS Amplicor HCMV Monitor for early detection and monitoring of human cytomegalovirus infection after allogeneic stem cell transplantation

Lengerke

Ljubicic

Meisner³

et al. 2006

Bone Marrow Transplant

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Early diagnosis of human cytomegalovirus (HCMV) infection and the introduction of preemptive antiviral therapy have reduced HCMV-related mortality after allogeneic stem cell transplantation. A critical goal remains stratifying risk profiles and minimizing potential harm owing to antiviral overtreatment. We compared the commercially available standardized COBAS Amplicor CMV Monitor (CACM) to an in-house PCR assay, for the monitoring of HCMV infection. Seventy-two patients were surveyed by an in-house PCR of whole blood, quantitative viral load assessment by CACM and virus culture assays in a prospective and a retrospective study. A high concordance between CACM and PCR was documented. The viral load at onset correlated with the peak viral load (Spearman rank correlation R ¼ 0.634, P ¼ 0.0004). In patients developing HCMV disease, both viral loads were in trend higher (P ¼ 0.823, respectively P ¼ 0.053), and the viremic episodes longer (P ¼ 0.015), as compared to asymptomatically HCMV-infected patients. The serological pre-transplant status was the major risk factor for the development of HCMV disease, showing highest risk for seropositive patients receiving a seronegative graft, whereas donor type (related or unrelated) and graft type (bone marrow or peripheral blood mobilized stem cells) did not have an influence. HCMV infection proved to be a risk factor for the development of non-viral opportunistic infections (P ¼ 0.002).

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Evaluation of the NucliSens CMV pp67 assay for detection and monitoring of human cytomegalovirus infection after allogeneic stem cell transplantation

Cited by 25 publications

References 23 publications

Assessment of CMV load in solid organ transplant recipients by pp65 antigenemia and real‐time quantitative DNA PCR assay: Correlation with pp67 RNA detection

Assessment of CMV load in solid organ transplant recipients by pp65 antigenemia and real‐time quantitative DNA PCR assay: Correlation with pp67 RNA detection

Prevention of Cytomegalovirus Disease in Recipients of Allogeneic Stem Cell Transplants

Evaluation of the COBAS Amplicor HCMV Monitor for early detection and monitoring of human cytomegalovirus infection after allogeneic stem cell transplantation

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