Detection of CMV DNA: Is EDTA whole blood superior to EDTA plasma?

Koidl, Christoph; Bozic, Michael; Marth, Egon; Kessler, Harald H.

doi:10.1016/j.jviromet.2008.08.012

Cited by 29 publications

(19 citation statements)

References 11 publications

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“…A growing availability and variety of highquality assays oVer increasing choices for the molecular diagnostic laboratory to introduce the optimal test system. QuantiWcation of CMV DNA in whole blood samples has been suggested as the most sensitive method for monitoring of CMV infection in immunosuppressed patients with whole blood being the currently accepted practice at our clinical center [5,[17][18][19][20][21][22][23]. In this study, suitability of the quantitative artus™ CMV RG PCR kit in conjunction with automated sample preparation on the QIAsymphony™ SP instrument for quantitation of CMV DNA was evaluated.…”

Section: Discussionmentioning

confidence: 99%

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Rapid quantitation of cytomegalovirus DNA in whole blood by a new molecular assay based on automated sample preparation and real-time PCR

Raggam¹,

Bozic

Salzer

et al. 2010

Med Microbiol Immunol

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Quantitation of cytomegalovirus (CMV) DNA in whole blood samples has gained significance in the routine diagnostic laboratory. In this study, the analytical performance of the artus™ CMV RG PCR kit in conjunction with automated sample preparation on the new QIAsymphony™ SP instrument was evaluated. Clinically referred samples were tested and results compared to those obtained with the routinely used molecular test system. Accuracy testing showed results to be within ±0.2 log(10) unit of the expected panel results. The assay was linear (R = 0.9972) from a lower quantification limit of 148 copies/ml to 1.3 × 10(7) copies/ml. The between-day imprecision CV was 8-63%, and the within-run imprecision CV was 16-61%. When 100 clinically referred samples were tested, results obtained with the new test system showed an acceptable concordance with those obtained with the routinely used easyMAG™ sample preparation and CMV HHV6,7,8 R-gene™ test system. Discrepant results were found with low-titer samples containing CMV DNA concentrations under the lower limit of quantification or within half a log unit above. The time to results was comparable for both test systems. The QIAsymphony™ sample preparation and artus™ CMV RG PCR test system allows for a rapid, sensitive, precise, and accurate high-throughput quantitation of CMV DNA in whole blood in the routine diagnostic laboratory.

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Section: Discussionmentioning

confidence: 99%

“…There is currently no consensus on the optimal blood compartment for routine molecular CMV DNA testing. In several studies, whole blood or plasma has been found to be superior to peripheral blood leukocytes [3,5,[17][18][19][20][21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Rapid quantitation of cytomegalovirus DNA in whole blood by a new molecular assay based on automated sample preparation and real-time PCR

Raggam¹,

Bozic

Salzer

et al. 2010

Med Microbiol Immunol

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“…Whole blood is easy to process because it does not require complex sample preparation compared to preparation of leukocyte subpopulations (126). While studies have shown good correlations in viral load values among the various compartments, significantly higher viral load levels have been detected in whole blood (91,126,128,130,131). Accordingly, many authorities have advocated the use of whole blood for CMV DNA detection due to its higher sensitivity and ability to detect low-copynumber viral reactivation.…”

Section: Nucleic Acid-based Methodsmentioning

confidence: 99%

Clinical Utility of Viral Load in Management of Cytomegalovirus Infection after Solid Organ Transplantation

2013

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SUMMARY The negative impact of cytomegalovirus (CMV) infection on transplant outcomes warrants efforts toward improving its prevention, diagnosis, and treatment. During the last 2 decades, significant breakthroughs in diagnostic virology have facilitated remarkable improvements in CMV disease management. During this period, CMV nucleic acid amplification testing (NAT) evolved to become one of the most commonly performed tests in clinical virology laboratories. NAT provides a means for rapid and sensitive diagnosis of CMV infection in transplant recipients. Viral quantification also introduced several principles of CMV disease management. Specifically, viral load has been utilized (i) for prognostication of CMV disease, (ii) to guide preemptive therapy, (iii) to assess the efficacy of antiviral treatment, (iv) to guide the duration of treatment, and (v) to indicate the risk of clinical relapse or antiviral drug resistance. However, there remain important limitations that require further optimization, including the interassay variability in viral load reporting, which has limited the generation of standardized viral load thresholds for various clinical indications. The recent introduction of an international reference standard should advance the major goal of uniform viral load reporting and interpretation. However, it has also become apparent that other aspects of NAT should be standardized, including sample selection, nucleic acid extraction, amplification, detection, and calibration, among others. This review article synthesizes the vast amount of information on CMV NAT and provides a timely review of the clinical utility of viral load testing in the management of CMV in solid organ transplant recipients. Current limitations are highlighted, and avenues for further research are suggested to optimize the clinical application of NAT in the management of CMV after transplantation.

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“…In these patients, prevention and management of CMV disease through monitoring of CMV DNA is of paramount importance. [11][12][13][14][15][16][17] …”

Section: Introductionmentioning

confidence: 99%

Extraction of viral nucleic acids: Comparison of five automated nucleic acid extraction platforms

Verheyen

Kaiser

Bozic

et al. 2012

Journal of Clinical Virology

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Detection of CMV DNA: Is EDTA whole blood superior to EDTA plasma?

Cited by 29 publications

References 11 publications

Rapid quantitation of cytomegalovirus DNA in whole blood by a new molecular assay based on automated sample preparation and real-time PCR

Rapid quantitation of cytomegalovirus DNA in whole blood by a new molecular assay based on automated sample preparation and real-time PCR

Clinical Utility of Viral Load in Management of Cytomegalovirus Infection after Solid Organ Transplantation

Extraction of viral nucleic acids: Comparison of five automated nucleic acid extraction platforms

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