Simultaneous Detection and Quantitation of Cytomegalovirus, Epstein-Barr Virus, and Adenovirus by Use of Real-Time PCR and Pooled Standards

Quantitative real-time PCR has become the most widely used preemptive approach for managing cytomegalovirus (CMV), Epstein-Barr virus (EBV), and adenovirus infections in immunosuppressed patients. These three assays are normally available as separate tests, each using five quantitation standards tha...

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Bibliographic Details
Published in:Journal of Clinical Microbiology 2009-03, Vol.47 (3), p.765-770
Main Authors: Gunson, R.N, Maclean, A.R, Shepherd, S.J, Carman, W.F
Format: Article
Language:English
Subjects:
Adenoviridae - genetics
Adenoviridae - isolation & purification
Adenovirus
Biological and medical sciences
Cytomegalovirus
Cytomegalovirus - genetics
Cytomegalovirus - isolation & purification
DNA Primers - genetics
Epstein-Barr virus
Fundamental and applied biological sciences. Psychology
Herpesvirus 4, Human - genetics
Herpesvirus 4, Human - isolation & purification
Humans
Microbiology
Miscellaneous
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - standards
Reference Standards
Reproducibility of Results
Sensitivity and Specificity
Virology
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Summary:Quantitative real-time PCR has become the most widely used preemptive approach for managing cytomegalovirus (CMV), Epstein-Barr virus (EBV), and adenovirus infections in immunosuppressed patients. These three assays are normally available as separate tests, each using five quantitation standards that are tested in duplicate. We have developed an adenovirus-CMV-EBV triplex assay that uses one set of five pooled quantitative standards, tested singly rather than in duplicate. This test demonstrated a sensitivity and an accuracy of quantitation equivalent to those of our previous single tests and was shown to be able to detect mixed infections with no loss in sensitivity. This assay is now in routine use in our laboratory and has considerably simplified the work flow of the laboratory, with a resultant improvement in sample turnaround time and significantly reduced costs.
ISSN:0095-1137
1098-660X
DOI:10.1128/JCM.01721-08