Parallel detection of five human herpes virus DNAs by a set of real-time polymerase chain reactions in a single run

Background: Human herpes viruses cause a spectrum of diseases that are usually self-limiting but can be reactive during immuno-suppression and may then lead to severe or even life-threatening diseases. The LightCycler technology allows rapid polymerase chain reaction (PCR) including product analysis...

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Bibliographic Details
Published in:Journal of clinical virology 2003, Vol.26 (1), p.85-93
Main Authors: Stöcher, Markus, Leb, Victoria, Bozic, Michael, Kessler, Harald H, Halwachs-Baumann, Gabriele, Landt, Olfert, Stekel, Herbert, Berg, Jörg
Format: Article
Language:English
Subjects:
Anticoagulants
Biological and medical sciences
Blood - virology
Blood Specimen Collection
Cerebrospinal Fluid - virology
Computer Systems
Cytomegalovirus (CMV)
Cytomegalovirus - genetics
DNA, Viral - analysis
DNA, Viral - blood
DNA, Viral - cerebrospinal fluid
Edetic Acid
Epstein-Barr virus (EBV)
Fundamental and applied biological sciences. Psychology
Herpes simplex virus (HSV)
Herpesviridae - genetics
Herpesvirus 1, Human - genetics
Herpesvirus 3, Human - genetics
Herpesvirus 4, Human - genetics
Humans
LightCycler
Microbiology
Polymerase Chain Reaction - instrumentation
Polymerase Chain Reaction - methods
Real-time polymerase chain reaction (PCR)
Sensitivity and Specificity
Species Specificity
Techniques used in virology
Varicella-zoster virus (VZV)
Virology
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Summary:Background: Human herpes viruses cause a spectrum of diseases that are usually self-limiting but can be reactive during immuno-suppression and may then lead to severe or even life-threatening diseases. The LightCycler technology allows rapid polymerase chain reaction (PCR) including product analysis within a closed system. This approach has been demonstrated to be suitable for routine diagnostic virus detection. Several LightCycler PCR assays have been established to the detection of human herpes viruses. The assays vary in their detection formats and PCR cycling protocols. So, they cannot be performed within a single LightCycler run. Objectives: Development of four LightCycler PCR assays for parallel detection of DNA derived from human cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), and varicella-zoster virus (VZV) in a single run. Study design: Primers and hybridization probes were tailored to suit one LightCycler PCR program. LightCycler PCRs were established, detection limits were determined, and clinical samples were evaluated. Results: With quantified herpes virus type specific DNA spiked into cerebrospinal fluid, serum or EDTA plasma the detection limits were found either at 500 or 250 viral DNA copies per ml depending on the virus DNA specific PCR and on the specimen type used. The applicability of the new LightCycler assays for routine molecular testing was evaluated by testing 96 clinical samples. Conclusion: The developed set of LightCycler PCRs permits parallel detection of CMV, EBV, HSV-1, HSV-2, and VZV in a single LightCycler run. The new molecular assays can easily be used to the rapid, simple, and convenient detection of herpes virus DNA in cerebrospinal fluid, serum and EDTA plasma in the routine diagnostic laboratory.
ISSN:1386-6532
1873-5967
DOI:10.1016/S1386-6532(02)00042-2