Non-invasive detection of malaria parasites in vitro and in Cameroonian adults

项目摘要 疟疾仍然是撒哈拉以南地区单一病原体致病和死亡的主要感染性原因。 尽管在过去二十年里取得了很大的进展，但非洲儿童仍然面临着巨大的挑战。虽然快速诊断测试的出现 作为一个重大进步，这些测试与金标准显微镜检查一样，也存在灵敏度不足的问题， 并需要血样进行检测。较新的分子方法提供了更高的灵敏度，但大多数需要 与显微镜检查相比，实验室基础设施水平明显更高，成本更高，执行时间更长 和快速诊断测试。光声流式细胞术(PAFC)是一种允许非 基于激光吸收后产生的光声信号对各种细胞类型的侵入性检测。 该设备目前正用于人类黑色素瘤试验，并已证明具有极好的安全性。我们 在实验室和动物中率先将其用于疟疾的非侵入性检测 模型，并已在使用便携式原型的早期人体研究中证明了安全性。作为后续行动，我们 建议严格评估这一便携式样机在体内的分析性能，以检测 喀麦隆成人中的单纯恶性疟原虫疟疾。PAFC的结果将与高度 灵敏和特异的分子分析，以及标准的诊断方法。重要的是，参与者将 进行纵向研究，并进行严格的安全评估。作为第二个目标，我们将 确定PAFC在体外和体外检测和区分人类疟疾种类的能力 使用来自感染者的储存血液、体外培养和显微镜载玻片。整体而言，建议的 研究的目的是证明快速、安全和高度敏感的PAFC用于非侵入性 在未怀孕的成年人中检测疟疾。数据将为我们小组在#年扩大研究奠定基础 儿童、孕妇、严重疟疾患者、无症状个体的筛查，以及其他 药效学研究。

Project Summary Malaria remains the leading infectious cause of morbidity and mortality by a single pathogen in sub-Saharan African children despite great progress in the past two decades. While the advent of rapid diagnostic tests has been a major stride, these tests suffer from similarly inadequate sensitivity as the gold standard, microscopy, and require a blood sample for testing. Newer molecular methods offer enhanced sensitivity, but most require significantly higher levels of laboratory infrastructure, cost more, and take longer to perform than microscopy and rapid diagnostic tests. Photoaccoustic flow cytometry (PAFC) is a technique that allows for the non- invasive detection of various cell types based on photoaccoustic signals generated following laser absorption. The device is being used currently in melanoma trials in humans and has demonstrated excellent safety. We have pioneered its use for the non-invasive detection of malaria in laboratory settings, as well as in animal models, and have demonstrated safety in early human studies using a portable prototype. As a follow-up, we propose to rigorously assess the in vivo analytical performance of this portable prototype for the detection of uncomplicated Plasmodium falciparum malaria in adults in Cameroon. PAFC results will be compared to highly sensitive and specific molecular assays, alongside standard diagnostic methods. Importantly, participants will be studied longitudinally, and rigorous safety assessments will be conducted. As a second aim, we will determine the ability of PAFC to detect and distinguish human malaria species in vitro and ex vivo through the use of stored blood from infected individuals, in vitro culture, and microscopy slides. Overall, the proposed studies aim to demonstrate the rapid, safe, and highly sensitive capability of PAFC for the non-invasive detection of malaria in non-pregnant adults. Data will lay the groundwork for expanded studies by our group in children, pregnant women, severe malaria patients, screening of asymptomatic individuals, and additional pharmacodynamic studies.