High Variability between Results of Different In-House Tests for Cytomegalovirus (CMV) Monitoring and a Standardized Quantitative Plasma CMV PCR Assay

Müller, Lutz von; Hampl, Walter; Hinz, J; Meisel, Helga; Reip, Angela; Engelmann, Elisabeth; Heilbronn, Regine; Гартнер, Барбара; Krämer, Oliver; Einsele, Hermann; Hebart, Holger; Ljubicic, T; Löffler, Jürgen; Mertens, Thomas

doi:10.1128/jcm.40.6.2285-2287.2002

Cited by 32 publications

(30 citation statements)

References 9 publications

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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%

“…Cultures were fixed with cold methanol for 10 min. 14 Forty-nine out of 72 patients studied were HCMV positive according to the PCR result from whole blood and were considered as having a viremic episode. In 40 of these 49 patients (81.6%), both assays gave a positive result, whereas nine patients tested positive by the in-house PCR were CACM negative (18.3%).…”

Section: Virus Culturementioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Virus Culturementioning

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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“…In the literature, CMV detection in plasma is mostly less sensitive than the assays in blood cells; 10 however, more sensitive inhouse CMV PCR assays in plasma were recently established 9,13 and automated NA isolation could also provide a more sensitive diagnosis of active CMV infection. 11 In our study, the sensitivity of CMM in plasma was significantly increased using automated NA isolation with COBAS AmpliPrep and Total Nucleic Acid Isolation kit than the standard manual NA isolation.…”

Section: Discussionmentioning

confidence: 99%

“…10 Briefly, blood cells were isolated by dextran sedimentation, sonified for 15 s at 70 W (Branson Sonifier; Danbury, CT, USA) and co-incubated with HEL. After 4 h, the medium was changed (minimal essential medium supplemented with 10% fetal calf serum, 1% penicillin/ streptomycin, 1% glutamine, 1% non-essential amino acids) and the cultures were incubated at 371C.…”

Section: Shell Vial Culturementioning

confidence: 99%

“…CMV monitoring in plasma might be easier to standardize and to perform compared with assays using isolated blood cells; [6][7][8][9] however, generally the assays using plasma seemed to be less sensitive. 10 Automated nucleic acid (NA) isolation systems are now available and may contribute to a better standardized and more sensitive diagnosis of active CMV infection. 11 The present study was aimed at comparing CMV monitoring in blood cells and plasma investigating patients after T-cell-depleted hematopoietic stem cell transplantation.…”

Section: Introductionmentioning

confidence: 99%

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CMV monitoring using blood cells and plasma: a comparison of apples with oranges?

Müller

Hinz

Bommer

et al. 2007

Bone Marrow Transplant

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Quantitative cytomegalovirus (CMV) monitoring is still far from being standardized between transplant centers. In the present study, we compared assays for quantitative CMV monitoring using blood cells and plasma. Four hundred and thirty-five consecutive samples from 29 patients with active CMV infection after allogeneic Tcell-depleted hemopoietic stem cell transplantation were tested in parallel using pp65 antigenemia and quantitative CMV polymerase chain reaction (PCR) in blood cells and plasma (COBAS AMPLICOR CMV MONITOR). Although only 142 (53.1%) of 253 positive samples were concordantly identified by all three assays, the number of positive samples detected by each assay was not different and the quantitative values were correlated, provided that nucleic acid (NA) in plasma was isolated by COBAS AmpliPrep and not by the manual protocol. Six (18%) of 34 episodes with active CMV infection were not detected using CMV PCR in plasma; whereas in times of white blood cell aplasia or blast crisis of leukemia, samples with active CMV infection in plasma could not be detected using blood cells. We conclude that CMV monitoring in whole blood could be favorable compared with assays using plasma or blood cells alone. Automated NA isolation could become an attractive tool for a more sensitive and better standardized molecular diagnostics.

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Severe graft rejection, increased immunosuppression, and active CMV infection in renal transplantation

Müller

Schliep

Storck³

et al. 2006

J. Med. Virol.

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Associations between active cytomegalovirus (CMV) infection, graft rejection, rejection therapy, and clinical signs and symptoms have been shown repeatedly. However, the causes and the sequence of events remain an area of debate. Two hundred twenty five patients with cadaveric renal transplant were included in the present study. Clinical signs and symptoms, and the development of active CMV infections were recorded during the first 3 months after renal transplantation. CMV monitoring by pp65-antigenemia was performed followed by preemptive antiviral therapy. Delayed graft function and severe graft rejection followed by anti T-cell antibody therapy was associated with the development of active CMV infection. In contrast, the induction therapy with anti-T-cell antibodies was not associated with more active CMV infections. Post-transplant morbidity determined by fever, pneumonia, and duration of hospital stay was increased significantly in patients with active CMV infection. However, in times of preemptive antiviral therapy an increased morbidity occurred in association with severe graft rejection and not with active CMV infection alone. In patients with renal transplantation and preemptive antiviral therapy, the morbidity was no more influenced by the CMV serostatus although the prevalence of active CMV infection was obviously different between CMV exposed (D+/R+,D+/R-, D-/R+) and unexposed (D-/R-) patients. Severe graft rejection and increased immunosuppression could stimulate cooperatively active CMV infections whereas immunosuppression alone may not be as effective. Prevention of severe graft rejection may be important to decrease early post-transplant morbidity and also the development of active CMV infections after renal transplantation.

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High Variability between Results of Different In-House Tests for Cytomegalovirus (CMV) Monitoring and a Standardized Quantitative Plasma CMV PCR Assay

Cited by 32 publications

References 9 publications

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

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

CMV monitoring using blood cells and plasma: a comparison of apples with oranges?

Severe graft rejection, increased immunosuppression, and active CMV infection in renal transplantation

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