Molecular Detection of Schistosome Infections with a Disposable Microfluidic Cassette.
Molecular Detection of Schistosome Infections with a Disposable Microfluidic Cassette.
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DOI：
10.1371/journal.pntd.0004318
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发表时间：
2015-12
影响因子：
3.8
通讯作者：
Greenberg RM
中科院分区：
文献类型：
--
作者：
Song J;Liu C;Bais S;Mauk MG;Bau HH;Greenberg RM
据估计，血吸虫等寄生虫以及丝虫病和土壤传播的线虫感染了全世界至少10亿人，对人类健康和经济发展造成了破坏性影响。感染动态的诊断和监测以及治疗效果几乎完全依赖于不准确、劳动密集型和不可靠的方法。这些缺点在大规模药物管理计划中被放大并具有更大的意义，在这些计划中，有效性的衡量取决于对治疗成功（或失败）、疾病传播率的变化和可能出现的耐药性的准确监测。在这里，我们采用等温分子分析，如环介导的等温扩增（LAMP），以一个简单的，手持的，定制的现场准备的微流控装置，允许敏感和特异性检测血吸虫感染小鼠的血清和血浆（用即时血浆分离器分离）中的无细胞核酸。使用我们的设备检测无细胞曼氏链球菌DNA，无需事先从血液中提取。我们的芯片具有很高的灵敏度（~2 × 10−17 g/μL），在感染后一周，即寄生虫产卵开始前几周，就能检测到曼氏梭菌DNA的阳性信号。这些结果表明，将等温扩增策略与我们的芯片结合，可能是一种快速、简单、低成本地诊断专利前和慢性血吸虫感染的策略，以及潜在的治疗效果监测。全世界有数亿人感染血吸虫，对人类健康和经济发展造成毁灭性影响。治疗和控制这些疾病的主要挑战之一是准确诊断感染和监测传播动态。目前的寄生虫学和血清学诊断检测往往缺乏敏感性、准确性和可靠性；更敏感和特异的分子方法需要在资源有限的环境中通常无法获得的基础设施。在这里，我们使用LAMP（一种等温扩增技术），在我们开发的简单、手持式、定制的、现场就绪的微流控装置中，检测了血吸虫感染小鼠血清和血浆中的无细胞寄生蠕虫DNA。早在感染一周后就可以检测到血吸虫DNA，这比蠕虫成熟并开始产卵要早得多。该装置体积小，完全独立，可一次性使用，易于适应快速简单的接触点（POC）感染检测。从样品采集到诊断的整个过程可在现场进行，无需任何实验室设备。
Parasitic helminths such as schistosomes, as well as filarial and soil-transmitted nematodes, are estimated to infect at least a billion people worldwide, with devastating impacts on human health and economic development. Diagnosis and monitoring of infection dynamics and efficacy of treatment depend almost entirely on methods that are inaccurate, labor-intensive, and unreliable. These shortcomings are amplified and take on added significance in mass drug administration programs, where measures of effectiveness depend on accurate monitoring of treatment success (or failure), changes in disease transmission rates, and emergence of possible drug resistance. Here, we adapt isothermal molecular assays such as loop-mediated isothermal amplification (LAMP) to a simple, hand-held, custom-made field-ready microfluidic device that allows sensitive and specific detection of schistosome cell-free nucleic acids in serum and plasma (separated with a point-of-care plasma separator) from Schistosoma mansoni-infected mice. Cell-free S. mansoni DNA was detected with our device without prior extraction from blood. Our chip exhibits high sensitivity (~2x10−17 g/μL), with a positive signal for S. mansoni DNA detectable as early as one week post infection, several weeks before parasite egg production commences. These results indicate that incorporation of isothermal amplification strategies with our chips could represent a strategy for rapid, simple, low-cost diagnosis of both pre-patent and chronic schistosome infections as well as potential monitoring of treatment efficacy. Schistosomes infect hundreds of millions of people worldwide, with devastating effects on human health and economic development. One of the major challenges in treatment and control of these diseases is the accurate diagnosis of infection and monitoring of transmission dynamics. Current parasitological and serological diagnostic tests often lack sensitivity, accuracy, and reliability; more sensitive and specific molecular methods require infrastructure typically unavailable in resource-limited settings. Here, we have detected cell-free parasitic helminth DNA in serum and plasma from schistosome-infected mice using LAMP, an isothermal amplification technique, in a simple, hand-held, custom-made, field-ready microfluidic device we have developed. Schistosome DNA is detectable as early as one week following infection, long before the worms have matured and begun producing eggs. The device is small, entirely self-contained and disposable, and is readily adaptable to rapid and simple point-of-contact (POC) detection of infection. The entire process from sample collection to diagnosis can be carried out on-site, without any laboratory equipment.