Rapid diagnosis of duck plagues virus infection by loop-mediated isothermal amplification.
Duck virus enteritis is a serious disease among farmed and free-living ducks (Anatidae) and a constant threat to the commercial duck industry in China. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed to rapidly detect and diagnose duck plague virus (DPV) in both farmed and wild waterfowl, and compared with polymerase chain reaction (PCR) method and real-time PCR method in accuracy, sensitivity and specificity. A set of four specific primers was successfully designed to recognize six distinct genomic sequences of UL6 protein from DPV, including one forward inner primer, one back inner primer and two outer primers. The optimum reaction temperature and time were verified to be 61.5 degrees C and 60 min, respectively. Comparative experiments showed that LAMP assay was a simple, rapid, accurate, sensitive and specific method for detecting DPV, and was superior to PCR assay in sensitivity and specificity for DNA amplification. In addition, challenge tests indicated the newly developed LAMP method was more sensitive for the diagnosis of DPV infection than virus isolation and PCR. LAMP assay would be a good alternative method for on-farm disease diagnosis.
Ji
Jun
J
College of Animal Science, South China Agricultural University, Guangzhou 510642, PR China.
Du
Li Qin
LQ
Xie
Qing Mei
QM
Cao
Yong Chang
YC
Zuo
Ke Jing
KJ
Xue
Chun Yi
CY
Ma
Jing Yun
JY
Chen
Feng
F
Bee
Ying Zuo
YZ
D018138:Alphaherpesvirinae / Q000302:isolation & purification*; D000818:Animals; D004372:Ducks; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms