London Centre for Nanotechnology, University College London, London, WC1H 0AH, UK.
Division of Medicine, University College London, London, WC1E 6BT, UK.
Sci Rep. 2023 Mar 27;13(1):4990. doi: 10.1038/s41598-023-31238-y.
Schistosomiasis is a major neglected tropical disease targeted for elimination as a public health issue by 2030, however there is an urgent need for more sensitive and specific diagnostic tests suitable to resource-limited settings. Here we developed CATSH, a CRISPR-assisted diagnostic test for Schistosoma haematobium, utilising recombinase polymerase amplification, Cas12a-targeted cleavage and portable real-time fluorescence detection. CATSH showed high analytical sensitivity, consistent detection of a single parasitic egg and specificity for urogenital Schistosoma species. Thanks to a novel CRISPR-compatible sample preparation developed using simulated urine samples containing parasitic eggs, CATSH had a sample-to-result within 2 h. The components of CATSH can be lyophilised, reducing cold chain dependence and widening access to lower and middle-income countries. This work presents a new application of CRISPR diagnostics for highly sensitive and specific detection of parasitic pathogens in remote areas and could have a significant impact on the elimination of neglected tropical diseases.
血吸虫病是一种主要的被忽视的热带病，目标是到 2030 年消除其对公共卫生的影响，但迫切需要更敏感和更特异的诊断检测方法，适用于资源有限的环境。在这里，我们开发了 CATSH，一种基于 CRISPR 的对埃及血吸虫的诊断检测方法，利用重组酶聚合酶扩增、Cas12a 靶向切割和便携式实时荧光检测。CATSH 显示出很高的分析灵敏度，能一致检测到单个寄生卵，并对泌尿生殖道血吸虫种具有特异性。由于使用含有寄生卵的模拟尿液样本开发了一种新颖的兼容 CRISPR 的样本制备方法，CATSH 可在 2 小时内得出结果。CATSH 的组件可以冻干，减少对冷链的依赖，扩大中低收入国家的获得途径。这项工作展示了 CRISPR 诊断在偏远地区对寄生虫病原体进行高灵敏度和特异性检测的新应用，可能对消除被忽视的热带病产生重大影响。
