Real-time PCR assay for universal detection and quantitation of all five species of fowl adenoviruses (FAdV-A to FAdV-E)

Journal of Virological Methods
Ayse GünesMichael Hess

Abstract

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.

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Citations

Apr 3, 2014·Avian Diseases·Ayse GünesMichael Hess
Oct 20, 2015·Molecular Therapy : the Journal of the American Society of Gene Therapy·Julio Alonso-PadillaAndrew H Baker
Oct 31, 2015·Avian Pathology : Journal of the W.V.P.A·Joanna Sajewicz-KrukowskaKatarzyna Domańska-Blicharz
Nov 28, 2015·Avian Pathology : Journal of the W.V.P.A·Pedro F Vera-HernándezAngel E Absalón
Oct 22, 2013·Veterinary Microbiology·Győző L KajánMária Benkő
Feb 7, 2015·Archives of Virology·Jowita Samanta NiczyporukElżbieta Samorek-Salamonowicz
Oct 23, 2016·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Huixin LiShengwang Liu
Apr 25, 2017·Avian Pathology : Journal of the W.V.P.A·Jianchang WangWanzhe Yuan
Aug 2, 2018·Archives of Virology·Jowita Samanta Niczyporuk, Hanna Czekaj
Feb 14, 2019·Virus Genes·Katarzyna Domańska-BlicharzJoanna Sajewicz-Krukowska
May 2, 2019·Avian Pathology : Journal of the W.V.P.A·Penelope A Steer-CopeAmir H Noormohammadi
Dec 20, 2017·The Journal of General Virology·Qi SuPeng Zhao
Dec 15, 2019·Applied Microbiology and Biotechnology·Qing PanXiaomei Wang
Feb 16, 2020·BMC Veterinary Research·Jowita Samanta NiczyporukKarolina Piekarska
Oct 20, 2018·Avian Diseases·A GarmynM Verlinden
Jun 5, 2021·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Aslı MeteBeate M Crossley

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