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Archives of Virology
Published in: Archives of Virology 4/2015

Open Access 01-04-2015 | Original Article

Application of cross-priming amplification (CPA) for detection of fowl adenovirus (FAdV) strains

Authors: Jowita Samanta Niczyporuk, Grzegorz Woźniakowski, Elżbieta Samorek-Salamonowicz

Published in: Archives of Virology | Issue 4/2015

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Abstract

Fowl adenoviruses (FAdVs) are widely distributed among chickens. Detection of FAdVs is mainly accomplished by virus isolation, serological assays, various polymerase chain reaction (PCR) assays, and loop-mediated isothermal amplification (LAMP). To increase the diagnostic capacity of currently applied techniques, cross-priming amplification (CPA) for the detection of the FAdV hexon gene was developed. The single CPA assay was optimised to detect all serotypes 1-8a-8b-11 representing the species Fowl aviadenovirus A-E. The optimal temperature and incubation time were determined to be 68 °C for 2 h. Using different incubation temperatures, it was possible to differentiate some FAdV serotypes. The results were recorded after addition of SYBR Green I® dye, which produced a greenish fluorescence under UV light. The CPA products separated by gel electrophoresis showed different “ladder-like” patterns for the different serotypes. The assay was specific for all serotypes of FAdV, and no cross-reactivity was observed with members of the genus Atadenovirus, duck atadenovirus A (egg drop syndrome virus EDS-76 [EDSV]) or control samples containing Marek’s disease virus (MDV), infectious laryngotracheitis virus (ILTV) or chicken anaemia virus (CAV). The results of the newly developed FAdV-CPA were compared with those of real-time PCR. The sensitivity of CPA was equal to that of real-time PCR and reached 10−2.0 TCID50, but the CPA method was more rapid and cheaper than the PCR systems. CPA is a highly specific, sensitive, efficient, and rapid tool for detection of all FAdV serotypes. This is the first report on the application of CPA for detection of FAdV strains.
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Metadata
Title
Application of cross-priming amplification (CPA) for detection of fowl adenovirus (FAdV) strains
Authors
Jowita Samanta Niczyporuk
Grzegorz Woźniakowski
Elżbieta Samorek-Salamonowicz
Publication date
01-04-2015
Publisher
Springer Vienna
Published in
Archives of Virology / Issue 4/2015
Print ISSN: 0304-8608
Electronic ISSN: 1432-8798
DOI
https://doi.org/10.1007/s00705-015-2355-9

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