Recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) designed for rapid detection of canine distemper virus

文献类型： 外文期刊

第一作者： Zhang, Shanshan

作者： Zhang, Shanshan;Wang, Chengyu;Meng, Keyin;Liu, Jun

作者机构：

关键词： canine distemper virus; lateral flow dipstick; recombinase polymerase amplification; sensitivity; specificity

期刊名称：JOURNAL OF VETERINARY MEDICAL SCIENCE （ 影响因子：1.1； 五年影响因子：1.1 ）

ISSN： 0916-7250

年卷期： 2024 年 86 卷 5 期

页码：

收录情况： SCI

摘要： In the present study, recombinase polymerase amplification (RPA) was combined with the colloidal gold lateral flow dipstick (LFD) method to establish a new, stable, and efficient assay for the detection of canine distemper virus (CDV). We designed a set of specific primers labeled with biotin and a specific probe labeled with dSpacer and C3 spacer, according to the conserved region in the N-terminal gene sequence of CDV. The reaction conditions and systems were then optimized, and the sensitivity and specificity were analyzed for potential clinical application. The results showed that the RPA-LFD assay for CDV detection was successfully established. We also found that the temperature in a closed fist (35 degrees C) is optimal for the RPA reaction. The optimal ratio of primer to probe was 2:1. The minimum detection limit of the RPA-LFD assay was 1 x 10(1) the median tissue culture infective dose (TCID50)/mL. Using this assay with samples from experimentally infected dogs, CDV was detected in nasal secretions, eye secretions, and blood on the fourth day post infection.In summary, this novel RPA-LFD assay for CDV detection is simple to use, and preliminary findings indicate its high specificity and sensitivity

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