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

被引:3
作者
Zhang, Shanshan [1 ]
Wang, Chengyu [1 ]
Meng, Keyin [1 ]
Liu, Jun [1 ]
机构
[1] Chinese Acad Agr Sci, Changchun Vet Res Inst, 666 Liuying West Rd, Changchun 130122, Peoples R China
关键词
canine distemper virus; lateral flow dipstick; recombinase polymerase amplification; sensitivity; specificity; NESTED PCR; RT-PCR; DOGS; DIAGNOSIS; BLOOD;
D O I
10.1292/jvms.23-0389
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
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
引用
收藏
页码:584 / 591
页数:8
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