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Detection and typin...
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Namvar, LillyGothenburg University,Göteborgs universitet,Institutionen för laboratoriemedicin, Avdelningen för klinisk virologi,Institute of Laboratory Medicine, Dept of Clinical Virology
(author)
Detection and typing of Herpes Simplex virus (HSV) in mucocutaneous samples by TaqMan PCR targeting a gB segment homologous for HSV types 1 and 2.
- Article/chapterEnglish2005
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Numbers
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LIBRIS-ID:oai:gup.ub.gu.se/56087
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https://gup.ub.gu.se/publication/56087URI
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https://doi.org/10.1128/JCM.43.5.2058-2064.2005DOI
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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Herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) are major causes of mucocutaneous lesions and severe infections of the central nervous system. Here a new semiautomated method for detecting and typing of HSV was used to analyze 479 mucocutaneous swab samples. After DNA extraction using a Magnapure LC robot, a 118-bp segment of the gB region was amplified by real-time PCR utilizing type-specific TaqMan probes to identify HSV-1 or HSV-2. HSV detection in a single well using probes labeled with carboxyfluorescein (FAM) for HSV-1 and JOE (6-carboxy-4',5'-dichloro-2',7'-dimethoxyfluorescein) for HSV-2 had a sensitivity similar to that seen in separate reactions. All but one of 217 samples (99.5%) that had been positive by virus culture were positive by TaqMan PCR, with a correct identification of type in all cases. Out of 262 samples negative by virus culture, 48 (18.3%) were positive by TaqMan PCR, with higher Ct values compared with culture positive samples (P < 0.0001). Overall, the Ct values for HSV-1 were lower than for HSV-2 (mean, 25.5 versus 27.9), but to some extent this could be due to weaker fluorescence by JOE. Lower C(t) values for HSV-1 were seen also in the 202 genital samples (79 HSV-1, 122 HSV-2, 1 HSV-1 and HSV-2), indicating that HSV-1 replicates as well as HSV-2 in the genital area. HSV-1 constituted 40% of genital infections and was associated with lower mean age (29.2 versus 36.4 years), probably reflecting the fact that recurrent genital HSV-1 infections are rare.
Subject headings and genre
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Adolescent
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Adult
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Aged
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Base Sequence
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DNA Primers
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DNA
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Viral
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genetics
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isolation & purification
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Herpes Simplex
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diagnosis
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Herpesvirus 1
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Human
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classification
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genetics
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isolation & purification
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Herpesvirus 2
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Human
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classification
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genetics
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isolation & purification
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Humans
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Middle Aged
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Polymerase Chain Reaction
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methods
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Reproducibility of Results
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Sensitivity and Specificity
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Taq Polymerase
Added entries (persons, corporate bodies, meetings, titles ...)
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Olofsson, Sigvard,1948Gothenburg University,Göteborgs universitet,Institutionen för laboratoriemedicin, Avdelningen för klinisk virologi,Institute of Laboratory Medicine, Dept of Clinical Virology(Swepub:gu)xolosi
(author)
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Bergström, Tomas,1950Gothenburg University,Göteborgs universitet,Institutionen för laboratoriemedicin, Avdelningen för klinisk virologi,Institute of Laboratory Medicine, Dept of Clinical Virology(Swepub:gu)xberto
(author)
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Lindh, Magnus,1960Gothenburg University,Göteborgs universitet,Institutionen för laboratoriemedicin, Avdelningen för klinisk virologi,Institute of Laboratory Medicine, Dept of Clinical Virology(Swepub:gu)xlmagr
(author)
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Göteborgs universitetInstitutionen för laboratoriemedicin, Avdelningen för klinisk virologi
(creator_code:org_t)
Related titles
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In:Journal of clinical microbiology43:5, s. 2058-640095-1137
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