title: [Real-time PCR as an efficient tool for investigating the presence of human herpesvirus 6 DNA].
abstract: Human herpesvirus 6 (HHV-6) is a beta-herpesvirus widely spread within a population and has been recognized as a potential significant pathogen in immunocompromised patients. Different clinical manifestations have been described including fever, skin rash, pneumonia, graft rejection and encephalitis. The goal of the study was development of real-time PCR assay for detection of human herpesvirus type 6 DNA in clinical samples, using primers targeting a conserved region of the viral DNA polymerase gene and a specific TaqMan hydrolyzing probe. The analytical sensitivity of assay was tested using serial dilutions of HHV-6 DNA in range between 10(0) and 10(-6). Thirty plasma samples taken from a group of adult recipients of allogeneic HSCT were tested for the presence of HHV-6 DNA in the LightCycler system. For comparison commercial quantitative MutaREAL HHV-6 kit (ALPCO) was used, according to the manufacturer's instructions. Both LightCycler assays, including in-house real-time PCR, detected HHV-6 DNA in 13 specimens. The conclusion is that developed TaqMan-based probes real-time PCR test is very reliable and valuable for detection of low-copy viremia in plasma samples. The high level of sensitivity and accuracy provided by the LightCycler instrument is favorable for the use of this method in the detection of HHV-6 DNA in clinical specimens.
authors.affiliation: Katedra i Zakład Mikrobiologii Lekarskiej, Warszawski Uniwersytet Medyczny. dzieciatkowski@wp.pl
authors.forename: Tomasz
authors.initials: T
authors.lastname: Dzieciatkowski
authors.forename: Maciej
authors.initials: M
authors.lastname: Przybylski
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Gieryńska
authors.forename: Mirosław
authors.initials: M
authors.lastname: Luczak
mesh_terms: D004279:DNA, Viral / Q000032:analysis*; D015654:Herpesvirus 6, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D014766:Viremia / Q000175:diagnosis* / Q000821:virology*
chemical_list: D004279:DNA, Viral