A Self-Contained Assay Device for Rapid Detection of Viable Cryptosporidium in Wa

一种用于快速检测WA中活隐孢子虫的独立检测装置

• 批准号: 8645032
• 负责人: XIAOLI SU
• 金额: $ 14.24万
• 依托单位: BIODETECTION INSTRUMENTS, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8645032
• 关键词: Address; Antibodies; Antigens; Biological Assay; Centers for Disease Control and Prevention (U.S.); Communities; Cryptosporidiosis; Cryptosporidium; Cryptosporidium parvum; Cyclospora; Cyst; Detection; Development; Devices; Disease; Disinfection; Dose; Enzymes; Evaluation; Giardia; Government; Growth; Immunoassay; In Vitro; Infection; Legal patent; Link; Marketing; Methods; Microscopic; Monitor; Oocysts; Parasites; Persons; Phase; Plants; Play; Process; Protocols documentation; Protozoa; Public Health; Research; Resistance; Risk; Role; Sampling; Small Business Innovation Research Grant; Sporozoites; Staging; System; Techniques; Technology; Testing; Time; United States; Water; Water Supply; base; chlorination; commercial application; cost effective; drinking water; excystation; human disease; improved; instrument; microbial; pathogen; prototype; public health relevance; rapid detection; rapid technique; transmission process; water sampling; water treatment; waterborne

DESCRIPTION (provided by applicant): This Small Business Innovation Research Phase I project will investigate the technical feasibility of an in vitro excystation-based automated cartridge assay for rapid detection of viable Cryptosporidium oocysts in water. Briefly, viable oocysts will be subjected to an in vitro excystation step to release sporozoites and the antigens for detection on a patent-pending self- contained cartridge immunoassay system. The Phase I research will focus on demonstrating the feasibility of specific detection of viable oocysts with a direct detection limit (i.e., without sample preconcentration) of d100-1000 oocysts/mL within in 1-2 h, which will be improved in Phase II to achieve the detection of d100 oocysts/10 L with sample preconcentration. The proposed assay will ultimately be highly automated and provide quantitative results for viable oocysts within 4 h from sample acquisition, largely reducing the complexity and lengthiness of current standard methods and allowing for near real-time monitoring of contamination as water leaves a drinking water treatment facility. While the United States has one of the safest water supplies in the world, our water still plays an important role i the transmission of human disease. Parasitic protozoa such as Cryptosporidium, Giardia, and Cyclospora are responsible for 21% of all waterborne illnesses. Cryptosporidium, in particular, presents unique challenges to community water systems since oocysts are resistant to standard disinfection processes (e.g., chlorination, ozonation), are small enough to pass through conventional water plant filters, and have a low infectious dose. The public health significance of dead Cryptosporidium oocysts is minimal; however, when the oocysts are infective, the risk to public health can be enormous. Unfortunately, Current EPA and conventional ELISA and PCR methods cannot determine the viability or infectivity of detected oocysts. In vitro excystation is commonly used to determine the viability of Cryptosporidium. However, in vitro excystation of Cryptosporidium is almost exclusively followed by microscopic examination to determine the number of excysted oocysts, which is tedious and time consuming and requires a sample purification step. If successfully developed, the proposed assay will address an unmet need in the market for rapid detection of viable Cryptosporidium in water. While the proposed research focuses on detection of viable Cryptosporidium only, the in vitro excystation assay principle would also be applicable to detection of other viable waterborne disease-causing protozoan parasites, e.g. Giardia and Cyclospora, significantly broadening its application and commercial appeal.

描述（由申请人提供）：这个小企业创新研究第一阶段项目将研究一种基于体外脱囊的自动化检测盒快速检测水中活隐孢子虫卵囊的技术可行性。简而言之，将活卵囊进行体外脱囊步骤以释放子孢子和抗原，用于在正在申请专利的自含式盒免疫测定系统上检测。第一阶段的研究将集中在证明可行性的具体检测活卵囊与 直接检测极限（即，在1-2 h内检测d100-1000个卵囊/mL），这将在第II阶段进行改进，以实现在样品预浓缩的情况下检测d100个卵囊/10 L。拟议的检测最终将高度自动化，并提供定量结果的活卵囊在4小时内从样品采集，大大降低了目前的标准方法的复杂性和冗长性，并允许近实时监测污染的水离开饮用水处理设施。虽然美国拥有世界上最安全的供水系统之一，但我们的水仍然在人类疾病的传播中发挥着重要作用。寄生原生动物，如隐孢子虫，贾第虫和环孢子虫是负责所有水传播疾病的21%。特别是隐孢子虫，对社区水系统提出了独特的挑战，因为卵囊对标准消毒过程有抗性（例如，氯化、臭氧化），小到足以通过常规的水厂过滤器，并且具有低的感染剂量。的公共卫生意义 死亡的隐孢子虫卵囊是最小的;然而，当卵囊具有传染性时，对公共健康的风险可能是巨大的。不幸的是，目前的EPA和传统的ELISA和PCR方法不能确定检测到的卵囊的活力或感染性。体外脱囊法常用于测定隐孢子虫的存活力。然而，在隐孢子虫的体外脱囊之后几乎完全是显微镜检查以确定脱囊卵囊的数量，这是繁琐和耗时的，并且需要样品纯化步骤。如果成功开发，拟议的检测方法将解决市场上快速检测水中活隐孢子虫的未满足需求。虽然拟议的研究只集中在检测活的隐孢子虫，体外脱囊测定原理也适用于检测其他活的水生致病原生动物寄生虫，如贾第虫和环孢子虫，显着扩大其应用和商业吸引力。