Rapid detection of group I avian adenoviruses by a loop-mediated isothermal amplification.
A loop-mediated isothermal amplification (LAMP) assay was optimized for the rapid detection of Group I avian adenoviruses. A set of six primers was designed from the DNA sequences of hexon genes from Group I avian adenovirus. The assay was performed in a water bath for 60 min at 63 C, and the amplification result was visualized by adding a fluorescence dye reagent or by inspecting the white sediment. The results showed that the LAMP assay could detect all 12 serotypes of Group I avian adenovirus and nine Guangxi Group I avian adenovirus isolates. This avian adenovirus Group I-specific LAMP assay could detect 238 copies of avian adenovirus. No cross-reactions were detected using the LAMP assay with avian adenoviruses type II and III or with other avian viruses. The ability of LAMP to detect Group I avian adenovirus isolates was further evaluated with 184 cloacal swab samples from poultry. In total, 72 out of 184 cloacal swab samples from poultry were identified as positive by LAMP, whereas 45 out of 184 were identified as positive by conventional PCR test. The Group I avian adenovirus specific LAMP results were further confirmed by real-time PCR. This specific LAMP method holds promise as a rapid and specific diagnostic assay for detection of samples from birds suspected of adenovirus infection.
Department of Biotechnology, Guangxi Veterinary Research Institute, 51 You Ai Road, Nanning, Guangxi 530001, China. xiezhixun@126.com
Zhixun
Z
Xie
Yi
Y
Tang
Qing
Q
Fan
Jiabo
J
Liu
Yaoshan
Y
Pang
Xianwen
X
Deng
Zhiqin
Z
Xie
Yi
Y
Peng
Liji
L
Xie
Mazhar I
MI
Khan
D000818:Animals; D000259:Aviadenovirus / Q000145:classification / Q000302:isolation & purification*; D002645:Chickens; D002681:China / Q000453:epidemiology; D004279:DNA, Viral / Q000145:classification / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D011201:Poultry Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
D004279:DNA, Viral