Development of an One-step Reverse Transcription Loop-mediated Isothermal Amplification Method for Rapid Detection of Bovine Viral Diarrhea Virus

This study aimed to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for detecting BVDV. Since gp48 gene of BVDV is among the most conserved regions, a set of four primers was designed to amplify six target sequences at the gp48 gene region for the RT-LAMP assa...

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Bibliographic Details
Published inHunan agricultural science & technology newsletter : HASTN Vol. 15; no. 10; p. 1826
Main Authors Yuan, Wanzhe, Wang, Teng, Wang, Jianchang, Li, Limin, Zhang, Xiuyuan, Sun, Jiguo
Format Journal Article
LanguageEnglish
Published Changsha Wu Chu (USA-China) Science and Culture Media Corporation 01.10.2014
Subjects
Design
Diarrhea
Optimization
Viruses
Beijing China
United States > US
China
Hebei China
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Summary:This study aimed to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for detecting BVDV. Since gp48 gene of BVDV is among the most conserved regions, a set of four primers was designed to amplify six target sequences at the gp48 gene region for the RT-LAMP assay. The optimization of the RT-LAMP reaction was performed by evaluating reaction temperature and reaction time. The RT-LAMP assay was successfully conducted at 56 degrees Celsius within 40 min under isothermal conditions, and the results could be detected as ladder-like bands using agarose gel electrophoresis. The RT-LAMP assay is highly sensitive and able to detect 3.74x10^sup 10^ copies/μI of BVDV RNA, as no cross-reaction was observed with other viruses. Overall, the newly established RT-LAMP assay indicates the potential application in both clinical diagnosis and field surveillance of BVDV.
ISSN:1009-4229