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）：该小型企业创新研究第一阶段项目将研究基于体外排泄的自动盒式检测的技术可行性，以快速检测水中的活隐孢子虫卵囊。简而言之，活卵囊将经历体外排泄步骤，以释放子孢子和抗原，用于在正在申请专利的自包含盒免疫测定系统上进行检测。第一阶段研究将侧重于证明用特定检测活卵囊的可行性 1-2小时内直接检测限（即不加样品预浓缩）为d100-1000个卵囊/mL，二期将进一步改进，实现样品预浓缩检测d100个卵囊/10L。所提出的测定最终将实现高度自动化，并在样品采集后 4 小时内提供活卵囊的定量结果，大大降低了当前标准方法的复杂性和冗长性，并允许在水离开饮用水处理设施时近乎实时地监测污染情况。尽管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。隐孢子虫、贾第鞭毛虫和环孢子虫等寄生原生动物占所有水传播疾病的 21%。尤其是隐孢子虫，它给社区供水系统带来了独特的挑战，因为卵囊对标准消毒过程（例如氯化、臭氧化）有抵抗力，足够小，可以通过传统的水厂过滤器，并且感染剂量低。公共卫生意义 死亡的隐孢子虫卵囊极少；然而，当卵囊具有感染性时，对公共健康的风险可能是巨大的。不幸的是，当前的 EPA 和传统的 ELISA 和 PCR 方法无法确定检测到的卵囊的活力或感染性。体外排泄通常用于确定隐孢子虫的活力。然而，隐孢子虫的体外排泄几乎完全是通过显微镜检查来确定排泄的卵囊的数量，这是繁琐且耗时的，并且需要样品纯化步骤。如果开发成功，所提出的检测方法将解决市场上尚未满足的快速检测水中活隐孢子虫的需求。虽然拟议的研究仅侧重于检测活的隐孢子虫，但体外排泄测定原理也适用于检测其他活的水传播致病原生动物寄生虫，例如。贾第鞭毛虫和环孢子虫，显着扩大了其应用和商业吸引力。