EBV viral load detection in clinical virology

Гартнер, Барбара; Preiksaitis, Jutta K.

doi:10.1016/j.jcv.2010.03.016

Cited by 80 publications

(63 citation statements)

References 98 publications

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“…V iral load testing has become a routine part of clinical care, particularly for immunocompromised patients (1)(2)(3). Such tests are used to diagnose disease, trigger preemptive therapy, and determine treatment responsiveness and endpoints.…”

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confidence: 99%

Comparative Performance of Reagents and Platforms for Quantitation of Cytomegalovirus DNA by Digital PCR

Hayden

Sam

et al. 2016

J Clin Microbiol

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A potential benefit of digital PCR is a reduction in result variability across assays and platforms. Three sets of PCR reagents were tested on two digital PCR systems (Bio-Rad and RainDance), using three different sets of PCR reagents for quantitation of cytomegalovirus (CMV). Both commercial quantitative viral standards and 16 patient samples (n ‫؍‬ 16) were tested. Quantitative accuracy (compared to nominal values) and variability were determined based on viral standard testing results. Quantitative correlation and variability were assessed with pairwise comparisons across all reagent-platform combinations for clinical plasma sample results. The three reagent sets, when used to assay quantitative standards on the Bio-Rad system, all showed a high degree of accuracy, low variability, and close agreement with one another. When used on the RainDance system, one of the three reagent sets appeared to have a much better correlation to nominal values than did the other two. Quantitative results for patient samples showed good correlation in most pairwise comparisons, with some showing poorer correlations when testing samples with low viral loads. Digital PCR is a robust method for measuring CMV viral load. Some degree of result variation may be seen, depending on platform and reagents used; this variation appears to be greater in samples with low viral load values.

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confidence: 99%

Comparative Performance of Reagents and Platforms for Quantitation of Cytomegalovirus DNA by Digital PCR

Hayden

Sam

et al. 2016

J Clin Microbiol

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“…With the rapid development of real-time quantitative PCR, the measurement of EBV DNA load (EDL) during these EBV-associated diseases has been largely implemented in clinical practice (3)(4)(5). The monitoring of EDL in blood is required for transplant recipients at risk of posttransplantation lymphoproliferative disorders (PTLDs) and could also be a surrogate marker for the adjustment of the immunosuppressive regimen in these patients (6,7).…”

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confidence: 99%

“…EDL measurement in plasma also appears to be a useful biomarker for the management of EBV-associated undifferentiated nasopharyngeal carcinoma (8). Although less clearly demonstrated, EDL measurements could also be helpful in other clinical situations such as severe or atypical infectious mononucleosis and other EBV-associated malignancies in immunosuppressed or immunocompetent patients (3,9).…”

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Multicenter Evaluation of Whole-Blood Epstein-Barr Viral Load Standardization Using the WHO International Standard

et al. 2016

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The first WHO international standard for Epstein-Barr virus (EBV) (WHO EBV standard) for nucleic acid amplification technology (NAT)-based assays was commercialized in January 2012 by the National Institute for Biological Standards and Control. In the study reported here, we compared whole-blood EBV DNA load (EDL) results from 12 French laboratories for seven samples (Quality Controls for Molecular Diagnostics 2013 proficiency panel) in order to determine whether expression in international units reduces interlaboratory variability in whole-blood EDLs. Each testing laboratory used a conversion factor to convert EDL results from copies per milliliter to international units per milliliter. This conversion factor was calculated from the WHO EBV standard according to the protocol described in this study (nine laboratories) or the recommendations of the PCR kit suppliers (three laboratories). The interlaboratory variability in whole-blood EDL results was reduced after standardization of the results using the WHO EBV standard. For the seven samples tested, standard deviations (SD) ranged from 0.41 to 0.55 when the results were expressed in log copies per milliliter, whereas the SD ranged from 0.17 to 0.32 when results were given in log international units per milliliter. Comparing the variance data (F test), we showed that the dispersion of whole-blood EDL results was significantly lower when they were expressed in log international units per milliliter (P < 0.001 for six of seven samples and P < 0.05 for one sample with a low mean EDL of 2.62 log IU/ml). This study showed that the use of the WHO EBV standard could improve the homogeneity of whole-blood EDL results between laboratories as well as the monitoring of patients at high risk of posttransplant lymphoproliferative disorders or other EBV-associated diseases. Primary Epstein-Barr virus (EBV) infection is the cause of the vast majority of cases of infectious mononucleosis and the subsequent lifelong persistence of EBV in the host, which, although mostly asymptomatic, can lead to the development of several lymphoid and epithelial cancers in immunosuppressed and immunocompetent individuals (1, 2).With the rapid development of real-time quantitative PCR, the measurement of EBV DNA load (EDL) during these EBV-associated diseases has been largely implemented in clinical practice (3-5). The monitoring of EDL in blood is required for transplant recipients at risk of posttransplantation lymphoproliferative disorders (PTLDs) and could also be a surrogate marker for the adjustment of the immunosuppressive regimen in these patients (6, 7). EDL measurement in plasma also appears to be a useful biomarker for the management of EBV-associated undifferentiated nasopharyngeal carcinoma (8). Although less clearly demonstrated, EDL measurements could also be helpful in other clinical situations such as severe or atypical infectious mononucleosis and other EBV-associated malignancies in immunosuppressed or immunocompetent patients (3, 9).Besides the debates on the clinical utility ...

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“…outine viral load measurements have become the standard of care for many patients, particularly those with severely compromised immune systems (1)(2)(3)(4)(5). However, despite their widespread clinical use, current testing strategies still have many limitations.…”

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Comparative Evaluation of Three Commercial Quantitative Cytomegalovirus Standards by Use of Digital and Real-Time PCR

Hayden

Sam

et al. 2015

J Clin Microbiol

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The recent development of the 1st WHO International Standard for human cytomegalovirus (CMV) and the introduction of commercially produced secondary standards have raised hopes of improved agreement among laboratories performing quantitative PCR for CMV. However, data to evaluate the trueness and uniformity of secondary standards and the consistency of results achieved when these materials are run on various assays are lacking. Three concentrations of each of the three commercially prepared secondary CMV standards were tested in quadruplicate by three real-time and two digital PCR methods. The mean results were compared in a pairwise fashion with nominal values provided by each manufacturer. The agreement of results among all methods for each sample and for like concentrations of each standard was also assessed. The relationship between the nominal values of standards and the measured values varied, depending upon the assay used and the manufacturer of the standards, with the degree of bias ranging from ؉0.6 to ؊1.0 log 10 IU/ml. The mean digital PCR result differed significantly among the secondary standards, as did the results of the real-time PCRs, particularly when plotted against nominal log 10 IU values. Commercially available quantitative secondary CMV standards produce variable results when tested by different real-time and digital PCR assays, with various magnitudes of bias compared to nominal values. These findings suggest that the use of such materials may not achieve the intended uniformity among laboratories measuring CMV viral load, as envisioned by adaptation of the WHO standard.R outine viral load measurements have become the standard of care for many patients, particularly those with severely compromised immune systems (1-5). However, despite their widespread clinical use, current testing strategies still have many limitations. Except for HIV and hepatitis B and C viruses, there has been little standardization of the testing process. Most methods are based on real-time PCR and have a high degree of result variability, particularly when testing among institutions is compared (6-9). The reasons for this variability are myriad. Real-time PCR is a dynamic process, with quantification based on normalization of the time to signal generation to a calibration curve that is in turn based on the use of calibration material with "known" values. Variations in any part of this complex procedure might theoretically affect result accuracy or precision. In fact, several factors have been shown to play a role (10); however, the most emphasis in the literature has been placed on the lack of universally accepted calibrators (11, 12). The lack of available international quantitative standards for many of the commonly tested viral analytes has led to the use of a wide variety of materials, intuitively reducing the agreement of results when common samples have been tested by different centers. It has been widely hoped that the development of such international reference material would help improve this situation. The...

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EBV viral load detection in clinical virology

Cited by 80 publications

References 98 publications

Comparative Performance of Reagents and Platforms for Quantitation of Cytomegalovirus DNA by Digital PCR

Comparative Performance of Reagents and Platforms for Quantitation of Cytomegalovirus DNA by Digital PCR

Multicenter Evaluation of Whole-Blood Epstein-Barr Viral Load Standardization Using the WHO International Standard

Comparative Evaluation of Three Commercial Quantitative Cytomegalovirus Standards by Use of Digital and Real-Time PCR

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