Application of Cross-priming amplification (CPA) for detection of fowl adenovirus (FAdV) strains
PBN-AR
Instytucja
Państwowy Instytut Weterynaryjny - Państwowy Instytut Badawczy
Informacje podstawowe
Główny język publikacji
en
Czasopismo
ARCHIVES OF VIROLOGY
ISSN
0304-8608
EISSN
1432-8798
Wydawca
SPRINGER WIEN
DOI
URL
Rok publikacji
2015
Numer zeszytu
4
Strony od-do
1005-1013
Numer tomu
160
Identyfikator DOI
Liczba arkuszy
Autorzy
Słowa kluczowe
angielski
Fowl adenoviruses
FAdVs
chickens
cross-priming amplification
CPA
Streszczenia
Język
angielski
Treść
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.
Cechy publikacji
ORIGINAL_ARTICLE
Inne
System-identifier
597099
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