Malaria is caused by parasites of the Plasmodium genus and remains one of the most pressing human health problems. The spread of parasites resistant to or partially resistant to single or multiple drugs, including frontline antimalarial artemisinin and its derivatives, poses a serious threat to current and future malaria control efforts. In vitro drug assays are important for identifying new antimalarial compounds and monitoring drug resistance. Due to its robustness and ease of use, the [³H]-hypoxanthine incorporation assay is still considered a gold standard and is widely applied, despite limited sensitivity and the dependence on radioactive material. Here, we present a first-of-its-kind chemiluminescence-based antimalarial drug screening assay. The effect of compounds on P. falciparum is monitored by using a dioxetane-based substrate (AquaSpark β-D-galactoside) that emits high-intensity luminescence upon removal of a protective group (β-D-galactoside) by a transgenic β-galactosidase reporter enzyme. This biosensor enables highly sensitive, robust, and cost-effective detection of asexual, intraerythrocytic P. falciparum parasites without the need for parasite enrichment, washing, or purification steps. We are convinced that the ultralow detection limit of less than 100 parasites of the presented biosensor system will become instrumental in malaria research, including but not limited to drug screening.

中文翻译：

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用于抗疟药物发现的下一代化学发光探针

疟疾是由疟原虫属寄生虫引起的，仍然是最紧迫的人类健康问题之一。对单一或多种药物（包括一线抗疟药物青蒿素及其衍生物）具有耐药性或部分耐药性的寄生虫的传播，对当前和未来的疟疾控制工作构成了严重威胁。体外药物测定对于识别新的抗疟化合物和监测耐药性非常重要。由于其稳健性和易用性，[ ³ H]-次黄嘌呤掺入测定仍然被认为是金标准并被广泛应用，尽管灵敏度有限并且依赖于放射性物质。在这里，我们提出了一种首创的基于化学发光的抗疟药物筛选方法。使用基于二氧杂环丁烷的底物（AquaSpark β-D-半乳糖苷）监测化合物对恶性疟原虫的影响，该底物在转基因 β-半乳糖苷酶去除保护基（β-D-半乳糖苷）后会发出高强度发光报告酶。该生物传感器能够高度灵敏、稳定且经济高效地检测无性、红细胞内恶性疟原虫寄生虫，无需寄生虫富集、洗涤或纯化步骤。我们相信，所提出的生物传感器系统对少于 100 种寄生虫的超低检测限将有助于疟疾研究，包括但不限于药物筛选。