Establishment of a nucleic acid dipstick test for detection of Schistosoma japonicum specific gene fragments based on the recombinase-aided isothermal amplification assay
10.16250/j.32.1374.2021016
- VernacularTitle:基于重组酶介导核酸等温扩增技术的日本血吸虫 特异性基因片段核酸试纸条检测方法的建立
- Author:
Yu-Ying YE
1
;
Song ZHAO
1
;
Yan-Hong LIU
2
;
Jian-Feng ZHANG
1
;
Chun-Rong XIONG
1
;
Qing-Jie YING
2
;
Kun YANG
1
Author Information
1. Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China
2. Jiangsu Qitian Gene Technology Co., Ltd., China
- Publication Type:Journal Article
- Keywords:
Schistosoma japonicum;
Nucleic acid dipstick;
Recombinase-aided isothermal amplification;
Nucleic acid detection;
SjG28 gene
- From:
Chinese Journal of Schistosomiasis Control
2021;33(4):334-338
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a rapid test for detection of Schistosoma japonicum specific gene fragments based on the recombinase-aided isothermal amplification assay (RAA) and nucleic acid dipstick test. Methods The S. japonicum SjG28 gene fragment was selected as the target gene fragment, and the primers and fluorescent probe were designed and synthesized. Then, a S. japonicum nucleic acid dipstick test was established. The sensitivity of this dipstick test was evaluated by detecting different copies of recombinant plasmids containing the S. japonicum SjG28 gene fragment and different concentrations of genomic DNA from adult worms of S. japonicum, and the specificity of the dipstick test was evaluated by detecting the genomic DNA from Clonorchis sinensis, S. mansoni, Ancylostoma duodenale, S. haematobium, Babesia and Paragonimus westermani. Results The S. japonicum nucleic acid dipstick test based on the S. japonicum SjG28 gene fragment showed the minimum detectable limit of 10 copies/μL of the recombinant plasmid containing the S. japonicum SjG28 gene fragment and the minimum detectable limit of 1 pg/μL of S. japonicum genomic DNA, and the dipstick assay tested negative for the genomic DNA from C. sinensis, S. mansoni, A. duodenale, S. haematobium, Babesia and P. westermani. Conclusion A rapid, simple, and visualized assay is established for detection of S. japonicum specific gene fragments based on RAA and nucleic acid dipstick test.
