A Point-of-Care Test for Rapid Diagnosis of Valley Fever

用于快速诊断山谷热的床旁检测 项目摘要 球孢子菌病（山谷热）是一种呼吸道疾病引起的吸入孢子从沙漠土壤- 寄生真菌，球孢子菌属（Coccidioides spp.）在流行地区，高达30%的社区获得性肺炎（CAP）是 由球孢子菌属引起。1山谷热模仿其他病毒和细菌呼吸道感染， 诊断的挑战。每年在临床实验室进行近300万次山谷热测试 在人类身上。通常，一管血液被送到临床实验室，以测试抗球孢子虫的抗体。 使用酶免疫测定（EIA）、免疫扩散（ID）和/或补体的真菌抗原制备 固定（CF）测试。从抽血到化验结果的时间可以是48小时至2周。SBIR提案 专注于快速抗体测试，可以在10分钟内进行，而病人在场。 根据以前出版物中的线索，我们的初步数据表明，CTS 1，一种几丁质酶，是 初级血清反应性免疫显性球虫抗原。因此，我们开发了一种快速测试， 使用重组CTS 1（rCTS 1）检测球孢子虫特异性IgG或IgM，不依赖于 测试（例如人或犬）。除大型兽医市场外，该测试的主要人类市场是 紧急护理诊所，急诊科和许多初级保健实践。 首次创新：尽管使用侧向流技术的快速抗体检测并不新鲜， VF测试提供足够的灵敏度和特异性，并检测IgM或IgG抗体， 这种真菌将极大地促进人类（和犬类）患者的临床护理。一个例子是在 紧急护理诊所，患者患有由球孢子菌属引起的社区获得性肺炎。可以 在他们就诊期间，通过简单的手指针刺滴血，使用我们的快速测试获得准确的诊断。 如果没有一个快速，敏感和具体的测试，紧急护理医生是不太可能下令室颤 因为结果在患者出院后最多4天返回给医生。 相反，患者通常会收到肺炎抗生素的处方-这是不适当的 球孢子菌病的治疗-并留下不正确/不完整诊断的紧急护理， 跟进我们快速10分钟测试的创新之处在于，它在10分钟内提供了一个临床可操作的答案。 使用当前的测试方法，一滴血只需几分钟，而不是2-4天。 第二次创新：球孢子菌的生长。需要经过认证的BSL 3实验室，训练有素的人员 以及昂贵且耗时的特殊设施;这一BSL 3努力不能被夸大。我们 创新性假设是rCTS 1可以作为抗体的主要抗原靶点， 球孢子菌病我们将测试我们的rCTS-1为基础的快速测试使用VF患者血清范围的性能 滴度为1：2至1：1024，同时还将其与现有商业检测进行比较，以显示等效性， FDA要求的。对照血清包括患有其他真菌、病毒和细菌感染的患者。 与唯一的商业上可获得的用于谷热的侧向流测试（Immy的索纳测试）相比，Cactus Bio的 快速检测更快（10分钟比30分钟），可定量，更准确，可以使用全血， 并且易于进行一致的高质量生产。

A Point-Of-Care Test for Rapid Diagnosis of Valley Fever PROJECT SUMMARY Coccidioidomycosis (Valley Fever) is a respiratory disease caused by inhalation of spores from the desert soil- dwelling fungus, Coccidioides spp. In endemic areas, up to 30% of community acquired pneumonia (CAP) is caused by Coccidioides spp.1 Valley Fever mimics other viral and bacterial respiratory infections and is challenging to diagnose. Nearly three million tests for Valley Fever are done in clinical laboratories each year in humans. Typically, a tube of blood is sent to a clinical laboratory to test for antibodies against a Coccidioidal fungal antigen preparation using Enzyme ImmunoAssay (EIA), ImmunoDiffusion (ID) and/or complement fixation (CF) tests. The time from blood draw to assay result may be 48 hours to 2 weeks. This SBIR proposal focuses on a rapid antibody test that can be performed in 10 minutes while the patient is present. Based on clues in previous publications2,3, our preliminary data demonstrate that CTS1, a chitinase, is the primary sero-reactive immunodominant Coccidioidal antigen. Therefore, we developed a rapid test that employs recombinant CTS1 (rCTS1) to detect Coccidioidal-specific IgG or IgM, independent of species being tested (e.g. human or canine). Excluding the large veterinary market, the primary human market for this test is Urgent Care Clinics, Emergency Departments and many primary care practices. First Innovation: Although rapid antibody tests using lateral flow technology are not new, a 10-minute rapid test for VF that provides sufficient sensitivity and specificity and detects either IgM or IgG antibodies against the fungus would dramatically advance clinical care for human (and canine) patients. One example is in the urgent care clinic where a patient with a community acquired pneumonia caused by Coccidioides spp. could receive an accurate diagnosis using our rapid test during their visit with a simple finger-stick drop of blood. Without the availability of a rapid, sensitive and specific test, an urgent care physician is unlikely to order a VF test because the result comes back to the physician up to 4 days after the patient has been discharged. Instead, patients usually receive a prescription for an antibiotic for pneumonia–which is an inappropriate treatment for coccidioidomycosis–and leave urgent care with an incorrect/incomplete diagnosis, potentially lost to follow up. The innovation of our rapid 10-minute test is that it provides a clinically actionable answer in 10 minutes with a single drop of blood instead of 2-4 days using current testing methods. Second Innovation: Growth of Coccidioides spp. requires a certified BSL3 laboratory, highly trained personnel and special facilities which are expensive and time-consuming; this BSL3 effort cannot be overstated. Our innovative hypothesis is that rCTS1 can serve as the primary antigen target for antibodies in patients with coccidioidomycosis. We will test the performance of our rCTS-1-based rapid test using VF patient sera ranging in titers from 1:2 to 1:1024 while also comparing it with existing commercial tests to show equivalence as required by the FDA. Control sera include patients with other fungal, viral and bacterial infections. Compared to the only commercially available lateral flow test for Valley Fever (Immy’s Sona test), Cactus Bio’s rapid test is faster (10 minutes compared to 30 minutes), quantifiable, more accurate, can use whole blood, and is amenable to consistent, high-quality production.