Evaluation of CMV viral load using TaqMan??? CMV quantitative PCR and comparison with CMV Antigenemia in heart and lung transplant recipients

Guiver, Malcolm; Fox, Andrew; Mutton, Ken; Mogulkoc, Nesrin; Egan, Jim

doi:10.1097/00007890-200106150-00021

Cited by 87 publications

(69 citation statements)

References 29 publications

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“…This difference in reproducibility is probably explained by the difference in copy number of the monitors, i.e., when using a monitor of a higher concentration, such as the 3.18-log 10 concentration used in the paper by Yun (23), a lower SD value is obtained. Such a decrease in SD at increasing genome numbers is statistically expected and has been shown for another qPCR assay (11). A run control at a low concentration facilitates the monitoring of the detection capacity of the PCR but has a higher uncertainty in measurement.…”

Section: Discussionsupporting

confidence: 55%

“…At present, there are insufficient data to define a clinically significant CMV concentration in plasma; it differs by type of patient group (7) and may vary over time in the same individual due to a changing immune status (16). It has been stated that copy numbers below 10 3 to 10 4 per ml of blood are rarely associated with CMV disease (1,11,12,21), but the increased use of preemptive antiviral treatment (10,15) has complicated the understanding of what is significant. Therefore, it is not evident what an optimal concentration of monitor control means.…”

Section: Discussionmentioning

confidence: 99%

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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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Section: Discussionsupporting

confidence: 55%

Section: Discussionmentioning

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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“…Briefly, DNA was extracted from 1 ml volume of blood using the Nucleon blood extraction kit BCC1 (Nucleon Biosciences, Glasgow, UK) and amplified using a previously described Taqman quantitative CMV PCR assay. 9 The limit of detection for this assay was defined as 500 CMV DNA genome copies/ml. CMV antigenemia was estimated using the immunoassay described previously.…”

Section: Graft Versus Host Disease Prophylaxismentioning

confidence: 99%

“…CMV antigenemia was estimated using the immunoassay described previously. 9 CMV treatment protocol Antiviral therapy was initiated in patients with two consecutive positive CMV PCR results and viral DNA copy numbers o1000 (log 3) or a single positive CMV PCR result and viral DNA copy number X1000 (log values 4log 3). Initial treatment comprised i.v.…”

Section: Graft Versus Host Disease Prophylaxismentioning

confidence: 99%

“…The assay is of particular value in neutropenic patients, with detection of CMV reported up to 8 days before a positive antigen test. 8,9 Preemptive therapy based on CMV RT-PCR therefore allows treatment to be initiated earlier, and so should afford reduced treatment duration and risk of CMV disease compared with antigenemia testing. 10 The current cohort reports a single institution experience of RT-PCR for the detection of CMV DNA, and the institution of preemptive therapy for CMV disease based on these results in more than 500 patients undergoing SCT.…”

Section: Introductionmentioning

confidence: 99%

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Active CMV disease does not always correlate with viral load detection

Ruell

Barnes

Mutton

et al. 2007

Bone Marrow Transplant

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Human Cytomegalovirus

Hodinka

2023

ClinMicroNow

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This chapter emphasizes the fundamental characteristics and clinical importance of cytomegalovirus (CMV) and focuses on the laboratory tests that can be used to identify the virus and to monitor and predict disease and susceptibility to antiviral therapy, and on the philosophy behind the interpretation and reporting of test results in various defined patient populations. Molecular and immunologic techniques, including whole‐genome sequencing directly from clinical specimens, have been used to study variation among CMV strains. CMV infections are common and usually asymptomatic in otherwise healthy children and adults; however, the incidence and spectrum of disease in newborns and in immunocompromised hosts establish this virus as an important human pathogen. Histopathologic testing, antigenemia assays, and qualitative and quantitative molecular methods have been routinely used for the direct detection of CMV from clinical samples. In the immunocompromised host, the use of direct detection methods to diagnose CMV disease is most beneficial for patient care.

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Evaluation of CMV viral load using TaqMan??? CMV quantitative PCR and comparison with CMV Antigenemia in heart and lung transplant recipients

Abstract: The TaqMan system enables a rapid high-throughput of samples. The TaqMan CMV QPCR can be used as an accurate and robust alternative to the antigenemia test to predict CMV disease and to monitor effectiveness of treatment.

Cited by 87 publications

References 29 publications

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

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

Active CMV disease does not always correlate with viral load detection

Human Cytomegalovirus

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