[Detection of human adenoviruses with real-time PCR assay using TaqMan fluorescent probes].
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.
Rola
Agnieszka
A
Wydział Rolnictwa i Biologii, Szkoła Główna Gospodarstwa Wiejskiego w Warszawie.
Przybylski
Maciej
M
Dzieciatkowski
Tomasz
T
Turowska
Agnieszka
A
Łuczak
Mirosław
M
D000258:Adenovirus Infections, Human / Q000175:diagnosis / Q000821:virology*; D000260:Adenoviruses, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics; D002460:Cell Line; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D005456:Fluorescent Dyes; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D019914:Taq Polymerase
D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; D005456:Fluorescent Dyes; C018315:hexon capsid protein, Adenovirus; D019914:Taq Polymerase