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[Detection of human adenoviruses with real-time PCR assay using TaqMan fluorescent probes].
- Source :
-
Medycyna doswiadczalna i mikrobiologia [Med Dosw Mikrobiol] 2007; Vol. 59 (4), pp. 371-7. - Publication Year :
- 2007
-
Abstract
- Infections with human adenoviruses are common worldwide and cause a wide range of signs and symptoms. Nowadays in current diagnostics procedures older virological methods, such virus isolation in a cell cultures, are replaced with molecular biology tests. The aim of the study was development of real-time PCR assay for detection of human adenoviruses. DNA isolated from A549 cell line infected with five different HAdV strains was used for development of a qualitative real-time PCR assay for detection of all human adenoviruses using primers targeting a conserved region of the hexon gene and a specific TaqMan probe. The analytical sensitivity of real-time PCR assay was tested using serial dilutions of HAdV7 DNA in range between 10(0) and 10(-6). For comparison typical end-point detected PCR for adenovirus detection with the same DNA dilutions was made. The sensitivity of novel method; was about thousand-fold higher than older one. The conclusion is that real-time PCR is very advisable in diagnostics of diseases caused with adenoviruses. The high level of sensitivity, specificity, accuracy, and rapidity provided by this assay are favorable for the use in the detection of adenoviral DNA in clinical specimens, especially from neuroinfections or immunocompromised hosts.
- Subjects :
- Adenovirus Infections, Human diagnosis
Adenoviruses, Human classification
Adenoviruses, Human genetics
Base Sequence
Capsid Proteins genetics
Cell Line
DNA Primers
DNA, Viral isolation & purification
Humans
Sensitivity and Specificity
Species Specificity
Adenovirus Infections, Human virology
Adenoviruses, Human isolation & purification
DNA, Viral analysis
Fluorescent Dyes
Polymerase Chain Reaction methods
Taq Polymerase
Subjects
Details
- Language :
- Polish
- ISSN :
- 0025-8601
- Volume :
- 59
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Medycyna doswiadczalna i mikrobiologia
- Publication Type :
- Academic Journal
- Accession number :
- 18416129