Real-time fluorescence loop-mediated isothermal amplification for the diagnosis of hemorrhagic enteritis virus.
Suspected cases of hemorrhagic enteritis associated with hemorrhagic enteritis virus (HEV) are becoming more frequent among yellow chickens in the Guangdong Province of China. In this study, we have developed a one-step, ecumenical, real-time fluorescence loop-mediated isothermal amplification (RealAmp) assay for the rapid diagnosis of HEV. The RealAmp assay was performed at 63°C and reduced the assay time to 15min, using a simple and portable device, the ESE-Quant Tube Scanner. The detection limit of DNA was 1fg/μl, and the detection was specific only to HEV. We also used nested PCR to evaluate the application of the RealAmp assay. The coincidence rate of the two methods was 100%. Our data indicated that the RealAmp assay provides a sensitive, specific, and user-friendly diagnostic tool for the identification and quantification of HEV for field diagnosis and in laboratory research.
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Xuemei
X
Liu
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Yuhao
Y
Li
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Chenggang
C
Xu
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Jianru
J
Qin
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Jianyong
J
Hao
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Min
M
Feng
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Liqiang
L
Tan
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Weixin
W
Jia
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China.
Ming
M
Liao
Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, The People's Republic of China. Electronic address: weishengcao@yahoo.com.
Weisheng
W
Cao
D000257:Adenoviridae Infections / Q000662:veterinary* / Q000821:virology; D000818:Animals; D002645:Chickens; D002681:China; D004751:Enteritis / Q000662:veterinary* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity; D046809:Siadenovirus / Q000302:isolation & purification*; D013696:Temperature; D013997:Time Factors; D014730:Veterinary Medicine / Q000379:methods; D014773:Virology / Q000379:methods*
Hemorrhagic enteritis virus; Nested PCR; RealAmp