title: Application of cross-priming amplification (CPA) for detection of fowl adenovirus (FAdV) strains.
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.
authors: Department of Poultry Viral Disease, National Veterinary Research Institute, Partyzantów 57 Avenue, 24-100, Pulawy, Poland, jowita.niczyporuk@piwet.pulawy.pl.
authors: Jowita Samanta
authors: JS
authors: Niczyporuk
authors: Grzegorz
authors: G
authors: Woźniakowski
authors: Elżbieta
authors: E
authors: Samorek-Salamonowicz
mesh_terms: D000257:Adenoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D000818:Animals; D000259:Aviadenovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D001483:Base Sequence; D002642:Chick Embryo; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010802:Phylogeny; D011201:Poultry Diseases / Q000175:diagnosis / Q000821:virology*
chemical_list: D017931:DNA Primers
references: Anal Bioanal Chem. 2014 May;406(13):3069-78
references: 24748469
references: Nucleic Acids Res. 2000 Jun 15;28(12):E63
references: 10871386
references: Avian Pathol. 2013 Aug;42(4):297-303
references: 23859215
references: J Vet Diagn Invest. 2010 Jan;22(1):61-6
references: 20093684
references: Avian Pathol. 2004 Apr;33(2):164-70
references: 15276983
references: J Virol Methods. 1998 Aug;73(2):211-7
references: 9766892
references: J Virol Methods. 2012 Aug;183(2):147-53
references: 22561984
references: Nat Protoc. 2008;3(5):877-82
references: 18451795
references: J Virol Methods. 1999 Aug;81(1-2):199-203
references: 10488779
references: J Clin Microbiol. 2012 Dec;50(12):3881-5
references: 22993179
references: Mol Cell Probes. 2010 Apr;24(2):104-6
references: 19835950
references: J Vet Med Sci. 2009 Sep;71(9):1239-42
references: 19801907
references: Arch Virol. 2014 Jun;159(6):1439-44
references: 24327092
references: Avian Pathol. 2001 Dec;30(6):655-60
references: 19184959
references: Mol Cell Probes. 2004 Oct;18(5):293-8
references: 15294316
references: J Clin Microbiol. 2009 Mar;47(3):845-7
references: 19116359
references: Can J Vet Res. 2008 Apr;72(3):236-41
references: 18505186
references: Avian Dis. 2011 Dec;55(4):575-9
references: 22312976
references: Avian Dis. 2001 Jan-Mar;45(1):268-75
references: 11332495
references: Lett Appl Microbiol. 2014 Aug;59(2):200-6
references: 24716722
references: Avian Pathol. 1991 Dec;20(4):597-606
references: 18680057
references: Can J Vet Res. 1999 Apr;63(2):124-8
references: 10369570
references: Avian Dis. 2012 Mar;56(1):73-81
references: 22545531
references: Avian Dis. 2011 Sep;55(3):462-7
references: 22017048
references: J Appl Microbiol. 2010 Mar;108(3):917-24
references: 19737344
references: J Virol Methods. 2010 Dec;170(1-2):147-54
references: 20869988
references: J Virol. 2004 Dec;78(23):13366-9
references: 15542689
references: Vet Res Commun. 2002 Oct;26(7):577-85
references: 12416873
references: Avian Pathol. 1977;6(3):189-217
references: 18770328
references: Can J Vet Res. 2007 Apr;71(2):98-102
references: 17479772
references: Avian Dis. 1999 Jan-Mar;43(1):98-105
references: 10216765
references: J Vet Med Sci. 2004 Dec;66(12):1561-6
references: 15644608
references: J Virol Methods. 2009 Dec;162(1-2):267-71
references: 19643144
references: Vet Microbiol. 2013 Dec 27;167(3-4):357-63
references: 24139719
references: Avian Dis. 2014 Mar;58(1):71-7
references: 24758116
references: J Virol Methods. 1983 Dec;7(5-6):327-35
references: 6330150
references: Avian Dis. 2011 Dec;55(4):719-23
references: 22313000
references: Acta Vet Hung. 2011 Jun;59(2):279-85
references: 21665581
references: J Vet Diagn Invest. 2006 Mar;18(2):162-7
references: 16617696
references: J Clin Microbiol. 2005 Apr;43(4):1646-50
references: 15814979