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Sci Rep. 2018 May 30;8(1):8393. doi: 10.1038/s41598-018-26717-6.

Loop-mediated Isothermal Amplification-Single Nucleotide Polymorphism Analysis for Detection and Differentiation of Wild-type and Vaccine Strains of Mink Enteritis Virus.

Lin P1,2, Wang H3, Cheng Y1,2, Song S1,2, Sun Y1,2, Zhang M1,2, Guo L1,2, Yi L1,2, Tong M1,2, Cao Z1,2, Li S1,2, Cheng S1,2, Wang J4,5.

Author information

1
Key Laboratory of Special Animal Epidemic Disease, Ministry of Agriculture, Changchun, 130112, People's Republic of China.
2
Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, People's Republic of China.
3
Shandong Sinder Technology Co., Ltd, Zhucheng, Shandong, 262204, People's Republic of China.
4
Key Laboratory of Special Animal Epidemic Disease, Ministry of Agriculture, Changchun, 130112, People's Republic of China. wangjianke@caas.cn.
5
Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, People's Republic of China. wangjianke@caas.cn.

Abstract

Broad coverage of mink enteritis virus (MEV) vaccination program in northeast of China has provided effective protection from mink viral enteritis. Nevertheless, MEV vaccine failures were reported due to continually evolving and changing virulence of field variants or wild-type MEV. In this study, a combined loop-mediated isothermal amplification (LAMP) and single nucleotide polymorphism (SNP) method, named LAMP-SNP assay, was developed for detection and differentiation of wild-type and vaccine strains of MEV. Four primers in MEV-VP2-LAMP were used to detect both wild-type and vaccine strains of MEV in our previous publication, and other four primers in LAMP-SNP were designed to amplify the NS1 gene in wild-type MEV and only used to detect wild-type viruses. The LAMP-SNP assay was performed in a water bath held at a constant temperature of 65 °C for 60 min. LAMP-SNP amplification can be judged by both electrophoresis and visual assessment with the unaided eyes. In comparison with virus isolation as the gold standard in testing 171 mink samples, the percentage of agreement and relative sensitivity and specificity of the LAMP-SNP assay were 97.1, 100%, and 94.0%, respectively. There were no cross-reactions with other mink viruses. The LAMP-SNP assay was found to be a rapid, reliable and low-cost method to differentiate MEV vaccine and field variant strains.

PMID:
29849073
PMCID:
PMC5976767
DOI:
10.1038/s41598-018-26717-6
[Indexed for MEDLINE]
Free PMC Article

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