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Real time PCR assay for universal detection and quantitation of all five species of fowl adenoviruses FAdV A to FAdV E     
Yazarlar (5)
Doç. Dr. Ayşe GÜNEŞ BAYIR Doç. Dr. Ayşe GÜNEŞ BAYIR
Türkiye
Ana Marek
Beatrice Grafl
Mıchael Hess
Evelyn Berger
Devamını Göster
Özet
The present study describes the development of a SYBR Green based real-time polymerase chain reaction (real-time PCR) for detection and quantitation of all fowl adenovirus (FAdV) species. Primers were designed based on conserved nucleotide sequences within the 52K gene. Ten-fold serial dilutions of a vector DNA were used as standard for quantitation. The real-time PCR had an efficiency of 98%, a regression squared value of 0.999 and showed a range of 6.73-6.73×10(8) copies of FAdV DNA per reaction. The assay was highly specific for FAdVs and an exact quantitation of all 5 FAdV species (FAdV-A to FAdV-E) could be demonstrated. It was shown, that twelve FAdV serotypes (FAdV-1 to 8a, and 8b to 11) were detectable and quantifiable. Other viral genomes as well as uninfected chicken embryo liver (CEL) cells did not produce positive signal. Cloacal swabs were taken during the animal experiment, which was performed with all FAdV species. Shedding of FAdVs was investigated in cell culture, by conventional PCR and by the developed real-time PCR. The real-time PCR was found more sensitive than cell culture and conventional PCR. Detection and quantitation of FAdVs in different type of samples was possible by the new real-time PCR.
Anahtar Kelimeler
Fowl adenoviruses | Real-time PCR | 52K gene | Sensitivity | Virus shedding | Quantitation
Makale Türü Özgün Makale
Makale Alt Türü SSCI, AHCI, SCI, SCI-Exp dergilerinde yayımlanan tam makale
Dergi Adı JOURNAL OF VIROLOGICAL METHODS
Dergi ISSN 0166-0934
Dergi Tarandığı Indeksler SCI-Expanded
Dergi Grubu Q4
Makale Dili İngilizce
Basım Tarihi 01-2012
Cilt No 83
Sayı 2
Sayfalar 147 / 153
Doi Numarası 10.1016/j.jviromet.2012.04.005
Makale Linki https://ac.els-cdn.com/S0166093412001322/1-s2.0-S0166093412001322-main.pdf?_tid=0487e23b-e36a-4374-b217-55662a94f8a1acdnat=1551359105_bb94a4b4011042e501ac02bf8f91a50c