A RT-LAMP Assay for Detection of Novel Duck Reovirus

doi:10.13560/j.cnki.biotech.bull.1985.2015.08.011

Biotechnology Bulletin ›› 2015, Vol. 31 ›› Issue (8): 71-75.doi: 10.13560/j.cnki.biotech.bull.1985.2015.08.011

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A RT-LAMP Assay for Detection of Novel Duck Reovirus

Yu Kexiang¹, Ma Xiuli¹, Han Hongyu^{1, 2}, Liu Cunxia¹, Li Yufeng¹, Huang Bing¹, Song Minxun¹

1. Key Laboratory of Poultry Disease Diagnose and Immune of Shandong Province,Institute of Poultry,Shandong Academy of Agricultural Sciences,Ji’nan 250023; 2. College of Animal Science ＆Veterinary Medicine,Shandong Agricultural University,Taian 271018

Received:2014-12-10 Online:2015-08-21 Published:2015-08-22

Abstract

Abstract: A one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting novel duck reovirus (NDRV) was established with 4 primers based on 6 conserved positions of the S3 gene. The process of assay was completed by using Bst DNA within 45 min at constant 63℃. RT-LAMP assay had solid specificity because no amplification was found with the samples of 6 other common duck diseases. The minimum detection limit of the RT-LAMP assay was 0.1 pg of viral RNA, which was 100 times of RT-PCR. The results of clinical application showed that the coincidence rate between the assay and the method of virus isolation and identification was 98%, and the requirement of instrument for the assay was relatively low. Therefore, the assay is a potential useful technique for NDRV detection in the field.

Key words: novel duck reovirus, S3 gene, one-step reverse transcription loop-mediated isothermal amplification, low requirement for instrument

Yu Kexiang, Ma Xiuli, Han Hongyu, Liu Cunxia, Li Yufeng, Huang Bing, Song Minxun. A RT-LAMP Assay for Detection of Novel Duck Reovirus[J]. Biotechnology Bulletin, 2015, 31(8): 71-75.

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A RT-LAMP Assay for Detection of Novel Duck Reovirus

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