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Improved real-time PCR assay for detection and quantification of all 54 known types of human adenoviruses in clinical samples.

Abstract

BACKGROUND
Detection and quantification of adenoviruses (AdVs) causing life-threatening complications are important abilities in recognition of infection and management of immunocompromised patients. Due to the rapid increase in the number of known AdV types, most commercial tests for detection and identification of AdVs are outdated.
MATERIAL/METHODS
We designed an improved, easier and faster real-time quantitative polymerase chain reaction (RQ-PCR) method for detection and quantification of 54 types of human AdVs. A wide validation effort was undertaken to ensure confidence in highly sensitive and specific detection of AdVs in compromised patients. The validation process included evaluation of the method's suitability and reliability for use in routine diagnostics.
RESULTS
Due to high sensitivity (9.2×10² copies/ml) and broad dynamic range (7 log) we are able to detect specific viral DNA in large amounts of cell-free body fluids. The new assay is characterized by high precision and low variation within and between individual virus tests (CV=0.036%, CV=1.29%), low bias error (4%) and no cross-reactivity with other pathogens.
CONCLUSIONS
The implementation of this new assay in clinical and laboratory practice provides a rapid, reliable and less laborious method for detection and monitoring of AdV replication in immunocompromised patients. Moreover, it offers the ability to distinguish between active and latent infection and assess treatment efficiency.

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  • Authors

    Bil-Lula I, De Franceschi N, Pawlik K, Woźniak M

    Source

    Medical science monitor : international medical journal of experimental and clinical research 18:6 2012 Jun pg BR221-8

    MeSH

    Adenoviruses, Human
    Adolescent
    Adult
    Biological Assay
    Child
    Child, Preschool
    DNA Primers
    DNA, Viral
    Female
    Genotype
    Humans
    Infant
    Male
    Real-Time Polymerase Chain Reaction
    Reference Values
    Reproducibility of Results
    Sensitivity and Specificity
    Young Adult

    Pub Type(s)

    Journal Article
    Research Support, Non-U.S. Gov't

    Language

    eng

    PubMed ID

    22648243