Establishment and evaluation of a novel DNA detection method based on recombinase-aided isothermal amplification assay for Giardia lamblia
10.16250/j.32.1374.2020035
- VernacularTitle:重组酶介导的蓝氏贾第鞭毛虫特异性等温核酸扩增方法的建立及评价
- Author:
Bi-Xian NI
1
;
Yan-Hong LIU
2
;
Xiang-Zhen XU
1
;
Xiao-Ting WANG
1
;
Xiao-Min WU
1
;
Qing-Jie YING
2
;
Jun CAO
1
;
Yang DAI
1
Author Information
1. Key Laboratory of National Health Commission of Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Public Health Research Center, Jiangnan University, Wuxi 214064, China
2. Jiangsu Qitian Gene Technology Co., Ltd., China
- Publication Type:Journal Article
- Keywords:
Giardia lamblia;
Nucleic acid detection;
Recombinase-aided isothermal amplification;
Fluorescent
- From:
Chinese Journal of Schistosomiasis Control
2020;32(4):345-349
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a novel nucleic acid assay for detection of Giardia lamblia based on the recombinase-aided isothermal amplification (RAA) assay, and evaluate its sensitivity and specificity for detection of G. lamblia. Methods The specific primer sequences and florescent probes were designed and synthesized based on the G. lamblia β-giardin gene as the target gene, and a fluorescent RAA assay was established. The recombinant plasmids at various copies (containing the β-giardin gene target sequence) and the genomic DNA of G. lamblia at various concentrations were used as templates for the fluorescent RAA assay to assess the sensitivity, and the genomic DNA from G. lamblia, Schistosoma japonicum, Clonorchis sinensis, Cryptosporidium parvum, Ascaris lumbricoides, Salmonella and Shigella was used as templates to assess the specificity of the fluorescent RAA assay. Results A novel fluorescent RAA assay was successfully established for detection of G. lamblia, which allowed the rapid and specific amplification of the target gene fragments at 39 ℃ within 20 min. The sensitivities of the fluorescent RAA assay were 102 copies/μL and 1 pg/μL for detection of the recombinant plasmid and G. lamblia genomic DNA, respectively, and the fluorescent RAA assay was negative for detection of the genomic DNA from S. japonicum, C. sinensis, C. parvum, A. lumbricoides, Salmonella and Shigella, which showed a high specificity. Conclusions A fluorescent RAA assay, which is simple, sensitive and specific, is successfully established for nucleic acid detection of G. lamblia.
