Establishment of reverse transcription loop-mediated isothermal amplification for rapid detection and differentiation of canine distemper virus infected and vaccinated animals

文献类型： 外文期刊

第一作者： Liu, Da-Fei

作者： Liu, Da-Fei;Chai, Hong-Liang;Yin, Xiu-Chen;Hua, Yu-Ping;Liu, Da-Fei;Liu, Chun-Guo;Tian, Jin;Zhang, Xiao-Zhan;Yang, Tian-Kuo;Zhang, Hong-Ying;Liu, Ming;Qu, Lian-Dong;Jiang, Yi-Tong

作者机构：

关键词： Canine distemper virus (CDV);Loop-mediated isothermal amplification (LAMP);Attenuated vaccine;Differential diagnosis

期刊名称：INFECTION GENETICS AND EVOLUTION （ 影响因子：3.342； 五年影响因子：3.188 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Although widespread vaccination against canine distemper virus (CDV) has been conducted for many decades, several canine distemper outbreaks in vaccinated animals have been reported frequently. In order to detect and differentiate the wild-type and vaccine strains of the CDV from the vaccinated animals, a novel reverse transcription loop-mediated isothermal amplification (RI-LAMP) method was developed. A set of four primers two internal and two external were designed to target the H gene for the specific detection of wild-type CDV variants. The CDV-H RT-LAMP assay rapidly amplified the target gene, within 60 min, using a water bath held at a constant temperature of 65 degrees C. The assay was 100-fold more sensitive than conventional RT-PCR, with a detection limit of 10(-1) TCID50 ml(-1). The system showed a preference for wild-type CDV, and exhibited less sensitivity to canine parvovirus, canine adenovirus type 1 and type 2, canine coronavirus, and canine parainfluenza virus. The assay was validated using 102 clinical samples obtained from vaccinated dog farms, and the results were comparable to a multiplex nested RT-PCR assay. The specific CDV-H RI-LAMP assay provides a simple, rapid, and sensitive tool for the detection of canines infected with wild-type CDV from canines vaccinated with attenuated vaccine. (C) 2015 Elsevier B.V. All rights reserved.

分类号： R1

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