TY - JOUR
T1 - Real-time fluorescence loop-mediated isothermal amplification for the diagnosis of hemorrhagic enteritis virus
AU - Liu, Xuemei
AU - Li, Yuhao
AU - Xu, Chenggang
AU - Qin, Jianru
AU - Hao, Jianyong
AU - Feng, Min
AU - Tan, Liqiang
AU - Jia, Weixin
AU - Liao, Ming
AU - Cao, Weisheng
PY - 2014/4/21
Y1 - 2014/4/21
N2 - 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 15. min, using a simple and portable device, the ESE-Quant Tube Scanner. The detection limit of DNA was 1. fg/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.
AB - 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 15. min, using a simple and portable device, the ESE-Quant Tube Scanner. The detection limit of DNA was 1. fg/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.
KW - Hemorrhagic enteritis virus
KW - Nested PCR
KW - RealAmp
UR - https://www.scopus.com/pages/publications/84896261149
U2 - 10.1016/j.virusres.2014.01.023
DO - 10.1016/j.virusres.2014.01.023
M3 - Article
C2 - 24487182
AN - SCOPUS:84896261149
SN - 0168-1702
VL - 183
SP - 50
EP - 55
JO - Virus Research
JF - Virus Research
ER -