title: Novel means of viral antigen identification: improved detection of avian influenza viruses by proximity ligation.
abstract: Recent outbreaks of avian influenza in different parts of the world have caused major economic losses for the poultry industry, affected wildlife seriously and present a significant threat even to human public health, due to the risk for zoonotic transmission. The ability to recognize avian influenza viruses (AIVs) early is of paramount importance to ensure that appropriate measures can be taken quickly to contain the outbreak. In this study, the performance of a proximity ligation assay (PLA) for the detection of AIV antigens in biological specimens was evaluated. It is shown that PLA: (i) as a novel principle of highly sensitive antigen detection is extending the arsenal of tools for the diagnosis of AIV; (ii) is very specific, nearly as sensitive as a commonly used reference real-time PCR assay, and four orders of magnitude more sensitive than a sandwich ELISA, utilizing the same antibody; (iii) avoids the necessity of nucleic acids extraction, which greatly facilitates high-throughput implementations; (iv) allows the use of inactivated samples, which safely can be transported from the field to diagnostic laboratories for further analysis. In summary, the results demonstrate that PLA is suited for rapid, accurate and early detection of AIV.
authors.affiliation: Rudbeck Laboratory, Department of Genetics and Pathology, Uppsala University, Dag Hammarskjölds väg 20, SE-75685 Uppsala, Sweden.
authors.forename: Joerg
authors.initials: J
authors.lastname: Schlingemann
authors.forename: Mikael
authors.initials: M
authors.lastname: Leijon
authors.forename: Alia
authors.initials: A
authors.lastname: Yacoub
authors.forename: Heidi
authors.initials: H
authors.lastname: Schlingemann
authors.forename: Siamak
authors.initials: S
authors.lastname: Zohari
authors.forename: Anna
authors.initials: A
authors.lastname: Matyi-Tóth
authors.forename: István
authors.initials: I
authors.lastname: Kiss
authors.forename: Göran
authors.initials: G
authors.lastname: Holmquist
authors.forename: Ann
authors.initials: A
authors.lastname: Nordengrahn
authors.forename: Ulf
authors.initials: U
authors.lastname: Landegren
authors.forename: Björn
authors.initials: B
authors.lastname: Ekström
authors.forename: Sándor
authors.initials: S
authors.lastname: Belák
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D009980:Influenza A virus / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry / Q000821:virology; D012680:Sensitivity and Specificity
chemical_list: D000956:Antigens, Viral