Monitoring of Human Cytomegalovirus Infections and Ganciclovir Treatment in Heart Transplant Recipients by Determination of Viremia, Antigenemia, and DNAemia

Gerna, Giuseppe; Zipeto, Donato; Parea, M.; Revello, Maria Grazia; Silini, Enrico Maria; Percivalle, Elena; Zavattoni, Maurizio; Grossi, Paolo; Milanesi, G.

doi:10.1093/infdis/164.3.488

Cited by 282 publications

(184 citation statements)

References 33 publications

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“…in nuclei of infected cell cultures [14], prior to the appearance of MIE protein p72. This discovery was the basis for the development of a new diagnostic assay, which is referred to as pp65-antigenemia, and has been found to be particularly useful for monitoring HCMV infections [15,16].…”

Section: Hcmv Leukocyte and Endothelial Cell Tropismmentioning

confidence: 99%

The pentameric complex of human Cytomegalovirus: cell tropism, virus dissemination, immune response and vaccine development

Gerna

Revello

Baldanti

et al. 2017

Journal of General Virology

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Between the 1980s and 1990s, three assays were developed for diagnosis of human cytomegalovirus (HCMV) infections: leuko (L)-antigenemia, L-viremia and L-DNAemia, detecting viral protein pp65, infectious virus and viral DNA, respectively, in circulating leukocytes Repeated initial attempts to reproduce the three assays in vitro using laboratory-adapted strains and infected cell cultures were consistently unsuccessful. Results were totally reversed when wild-type HCMV strains were used to infect either fibroblasts or endothelial cells. Careful analysis and sequencing of plaque-purified viruses from recent clinical isolates drew attention to the ULb¢ region of the HCMV genome. Using bacterial artificial chromosome technology, it was shown by both gain-of-function and loss-of-function experiments that UL131-128 genes are indispensable for virus growth in endothelial cells and virus transfer to leukocytes. In addition, a number of clinical isolates passaged in human fibroblasts had lost both properties (leuko-tropism and endothelial cell-tropism) when displaying a mutation in the UL131-128 locus

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Section: Hcmv Leukocyte and Endothelial Cell Tropismmentioning

confidence: 99%

The pentameric complex of human Cytomegalovirus: cell tropism, virus dissemination, immune response and vaccine development

Gerna

Revello

Baldanti

et al. 2017

Journal of General Virology

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“…Previous reports have shown that PCR testing of whole blood is more sensitive than conventional cell culture [12,[14][15][16][17][18] and DEAFF [12,14,15,[18][19][20][21]. Zipeto et al [12] found that one-third of 293 blood samples taken from a mixed population of transplant recipients, patients with AIDS and neonates gave positive results in PCR tests, but negative results by DEAFF and culture.…”

Section: Discussionmentioning

confidence: 99%

Comparison of three PCR techniques for detecting cytomegalovirus (CMV) DNA in serum, detection of early antigen fluorescent foci and culture for the diagnosis of CMV infection

Evans

Gray

Wreghitt

et al. 1999

Journal of Medical Microbiology

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Three PCR assays were developed for detection of cytomegalovirus (CMV) DNA in serum and were evaluated with samples from organ transplant recipients. The Qiamp Blood Kit was efficient for extraction of DNA from sera. Single-round PCR of a 293-bp region of CMV DNA was sensitive and highly specific for CMV targets and was more sensitive than detection of early antigen fluorescent foci (DEAFF) testing or isolation of CMV from buffy coat by cell culture. The results of a significant proportion of buffy coat samples were not interpretable because of toxicity in conventional culture or DEAFF tests. A non-competitve quantitative PCR test and semi-quantitative PCR test for the detection of CMV DNA in serum yielded comparable results for samples taken serially from three bone marrow transplant recipients. Single-round PCR was superior to conventional techniques for the diagnosis of CMV infection, was simple to perform and was completed rapidly. The semi-quantitative technique has added advantages where quantification is important.

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“…Quantitative assays can be based on pp65 antigenemia (6,9), PCR with detection by an enzyme immunoassay (2,8,18,25), or other detection techniques (19,20). The introduction of a commercial quantitative PCR assay, the COBAS Amplicor CMV Monitor test (Roche Molecular Systems, Pleasanton, Calif.), has provided a system that is easily used for clinical routine diagnostics and enables comparisons between laboratories (First EU-QCCA human cytomegalovirus proficiency panel, Quality Control for Molecular Diagnostics [http://www .qcmd.org]).…”

mentioning

confidence: 99%

Comparison of a Duplex Quantitative Real-Time PCR Assay and the COBAS Amplicor CMV Monitor Test for Detection of Cytomegalovirus

Herrmann¹,

Larsson²,

Rubin³

et al. 2004

J Clin Microbiol

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A duplex quantitative real-time PCR (qPCR) assay was designed to detect both the polymerase gene (pol) and the glycoprotein gene (gB) of cytomegalovirus (CMV). The detection limit of the qPCR was determined to be 1 to 3 copies/reaction and the linear measure interval was 10 3 to 10 8 copies/ml. The qPCR system was compared to the COBAS Amplicor CMV Monitor test (COBAS) by an analysis of 138 plasma samples. Both systems detected CMV in 71 cases and had negative results for 33 samples. In addition, 34 samples were positive by qPCR and negative by the COBAS assay, but in no case was the COBAS result positive and the qPCR result negative. Thus, qPCR detected 48% more positive cases than the COBAS method. For samples with >10 5 copies/ml by qPCR, a saturation effect was seen in the COBAS assay and quantification required dilution. Copy numbers for pol and gB by qPCR generally agreed. However, the reproducibility of qPCR assays and the need for an international standard are discussed. Discrepant copy numbers for pol and gB by qPCR were found for samples from two patients, and sequence analysis revealed that the corresponding CMV strains were mismatched at four nucleotide positions compared with the gB fragment primer sequences. In conclusion, a duplex qPCR assay in a real-time format facilitates quantitative measurements and minimizes the risk of false-negative results.Human cytomegalovirus (CMV) is a common opportunistic pathogen found in immunocompromised patients, e.g., patients in a posttransplantation state or those infected with human immunodeficiency virus. Early detection and close monitoring of CMV infections are important for preventing the reactivation of endogenous CMV and for the surveillance of recipients of seropositive donor transplants. Appropriate antiviral therapy is based on a knowledge of the viral load as well as a quantification of the initial amount of CMV and the rate of increase between the last negative and the first positive PCR sample. These are independent risk factors for CMV disease

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Monitoring of Human Cytomegalovirus Infections and Ganciclovir Treatment in Heart Transplant Recipients by Determination of Viremia, Antigenemia, and DNAemia

Cited by 282 publications

References 33 publications

The pentameric complex of human Cytomegalovirus: cell tropism, virus dissemination, immune response and vaccine development

The pentameric complex of human Cytomegalovirus: cell tropism, virus dissemination, immune response and vaccine development

Comparison of three PCR techniques for detecting cytomegalovirus (CMV) DNA in serum, detection of early antigen fluorescent foci and culture for the diagnosis of CMV infection

Comparison of a Duplex Quantitative Real-Time PCR Assay and the COBAS Amplicor CMV Monitor Test for Detection of Cytomegalovirus

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