文献类型： 外文期刊
作者： Li, Xin 1 ; Dang, Zhisheng 2 ; Tang, Wenqiang 3 ; Zhang, Haoji 1 ; Shao, Jianwei 1 ; Jiang, Rui 5 ; Zhang, Xu 1 ; Huang, Fuqiang 1 ;
作者机构： 1.Foshan Univ, Sch Life Sci & Engn, Foshan 528225, Peoples R China
2.World Hlth Org WHO Collaborating Ctr Trop Dis, Natl Hlth Commiss Peoples Republ China NHC,Natl Ct, Natl Inst Parasit Dis,Chinese Ctr Dis Control & Pr, Key Lab Parasite & Vector Biol,Chinese Ctr Trop Di, Shanghai 200025, Peoples R China
3.State Key Lab Hulless Barley & Yak Germplasm Resou, Lhasa 850002, Peoples R China
4.Tibet Acad Agr & Anim Husb Sci, Lhasa 850002, Peoples R China
5.Huazhong Agr Univ, Coll Anim Sci & Vet Med, Wuhan 430070, Peoples R China
关键词： detection; CRISPR; Cas12a; suboptimal crRNA; light-activated crRNA; tandem repeats; POCT
期刊名称：BIOSENSORS-BASEL （ 影响因子：5.4； 五年影响因子：5.7 ）
ISSN：
年卷期： 2024 年 14 卷 3 期
页码：
收录情况： SCI
摘要： The rapid and accurate identification of parasites is crucial for prompt therapeutic intervention in parasitosis and effective epidemiological surveillance. For accurate and effective clinical diagnosis, it is imperative to develop a nucleic-acid-based diagnostic tool that combines the sensitivity and specificity of nucleic acid amplification tests (NAATs) with the speed, cost-effectiveness, and convenience of isothermal amplification methods. A new nucleic acid detection method, utilizing the clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nuclease, holds promise in point-of-care testing (POCT). CRISPR/Cas12a is presently employed for the detection of Plasmodium falciparum, Toxoplasma gondii, Schistosoma haematobium, and other parasites in blood, urine, or feces. Compared to traditional assays, the CRISPR assay has demonstrated notable advantages, including comparable sensitivity and specificity, simple observation of reaction results, easy and stable transportation conditions, and low equipment dependence. However, a common issue arises as both amplification and cis-cleavage compete in one-pot assays, leading to an extended reaction time. The use of suboptimal crRNA, light-activated crRNA, and spatial separation can potentially weaken or entirely eliminate the competition between amplification and cis-cleavage. This could lead to enhanced sensitivity and reduced reaction times in one-pot assays. Nevertheless, higher costs and complex pre-test genome extraction have hindered the popularization of CRISPR/Cas12a in POCT.
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