Medizinische Universität Graz - Research portal

Go directly to the nagivation.
Go directly to the content.

Navigation/Search

Selected Publication:

SHR Neuro Cancer Cardio Lipid Metab Microb

Stöcher, M; Hölzl, G; Stekel, H; Berg, J.
Automated detection of five human herpes virus DNAs by a set of LightCycler PCRs complemented with a single multiple internal control.
J Clin Virol. 2004; 29(3):171-178 Doi: 10.1016/S1386-6532(03)00121-5
Web of Science PubMed FullText FullText_MUG

Leading authors Med Uni Graz: Berg Jörg
Altmetrics:
Dimensions Citations:
Plum Analytics:
Scite (citation analytics):
Abstract:: BACKGROUND: Herpes viruses represent important causes of morbidity and mortality especially in immuno-compromised patients. To assist in rapid diagnosis real-time PCR assays have been developed for the detection of herpes virus DNA in patient specimens. A recently described set of real-time PCR assays using LightCycler technology enabled parallel detection of DNA from cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus type 1 and 2 (HSV-1/-2), and varicella-zoster virus (VZV) by using a single LightCycler program [J. Clin. Virol. 26 (2003) 85]. The set of assays lacked automation of DNA purification and of PCR mixture preparation, and was not furnished with measures to monitor for sample adequacy. OBJECTIVES: Development of a set of automated LightCycler-PCR assays for the detection CMV-, EBV-, HSV-1/-2- and VZV-DNA in plasma samples and complementation of the assays with internal amplification controls (ICs). STUDY DESIGN: The MagNA Pure LC instrument was used for automated DNA purification and automated preparation of PCR mixtures. A single multiple IC-DNA specific for all four herpes virus type-specific PCRs was generated and used in all four LightCycler assays. Detection limits were determined and clinical samples were evaluated. RESULTS: With quantified herpes virus type-specific reference DNA spiked into EDTA plasma, the detection limits were found at 250 copies/ml of CMV-, EBV-, HSV-1/-2-DNA and at 500 copies/ml of VZV-DNA. The novel set of assays was evaluated by testing 112 EDTA plasma samples. The use of the IC led to the detection of PCR-inhibited samples. CONCLUSION: The set of automated LightCycler assays was found rapid, markedly labour saving and suitable for the routine diagnostic laboratory. The use of the one internal control molecule simplified the assay protocol and allowed monitoring for sample adequacy.
Find related publications in this database (using NLM MeSH Indexing): Automation - instrumentation; Cytomegalovirus - genetics Cytomegalovirus - isolation and purification; DNA, Viral - blood; Herpesviridae - genetics Herpesviridae - isolation and purification; Herpesviridae Infections - virology; Herpesvirus 1, Human - genetics Herpesvirus 1, Human - isolation and purification; Herpesvirus 2, Human - genetics Herpesvirus 2, Human - isolation and purification; Herpesvirus 3, Human - genetics Herpesvirus 3, Human - isolation and purification; Herpesvirus 4, Human - genetics Herpesvirus 4, Human - isolation and purification; Humans -; Polymerase Chain Reaction - instrumentation Polymerase Chain Reaction - methods; Reference Standards -
Find related publications in this database (Keywords): cytomegalovirus (CMV); Epstein-Barr virus (EBV); herpes simplex virus (HSV); varicella-zoster virus (VZV); real-time PCR; internal control; MagnaPure LC; LightCycler