Serological Biomarkers for Coccidioidomycosis

项目总结 截至2017年，球孢子菌病(Cocci)也称为谷热(VF)被指定为须报告的 疾病在22个州(1)。美国大多数病例发生在亚利桑那州南部和加利福尼亚州，那里的 随着天真的个人，特别是老年退休人员的涌入，人口正在增加。由于球孢子虫Coccidioids sp. 在完全具有免疫能力的宿主中造成显著的发病率和死亡率，这是对 这些地区居住着美国10%的人口，不包括大量的游客或兼职人员 亚利桑那州和加利福尼亚州的常住人口。重要的是，报告标准要求实验室呈阳性。 检测包含性，但约50%的患者--他们最终会检测为阳性--在急性疾病期间检测为阴性(2)。测试 敏感度和特异度低，使病人和他们的医疗保健提供者没有准确的 诊断。因此，迫切需要更好的诊断方法来正确识别Cocci患者 确保适当的随访和治疗随之而来。 球藻(Coccidioidesspp.)在沙漠土壤中以菌丝体的形式生长，并产生孢子(节孢子)以在 不利生长条件的次数。当土壤受到干扰时，节孢子虫被气雾化并被吸入 引发感染。在寄主内部，孢子转化为含有内孢子的球体，内孢子生长在 阿龙。当球体破裂时，内生孢子被释放出来，每个孢子都可以扩散形成一个新的球体。 临床诊断很困难，因为患者的症状类似于其他细菌和病毒肺炎。 实验室诊断通常仅依靠血清学，但约50%的患者在检测时不呈阳性 病情严重，导致诊断不准确，治疗不当。我们建议现在是时候重新-- 血清反应性球虫抗原的评价。在过去的60年里，血清学诊断 球孢子菌病依赖于对试管沉淀素(TP)的IgM反应和对补体的IgG反应 固定(CF)抗原。因为这些抗原主要在菌丝体中表达，而不是在球体中表达 在宿主中生长的真菌形式，难怪许多患者血清阴性，因为他们没有 但在急性肺部疾病期间产生了针对球状抗原的抗体。 这项提议的目标是识别与急性感染患者发生反应的新的球虫抗原 塞拉。目的是利用最近发表的球孢子虫。蛋白质组创造一种核酸 可编程蛋白质阵列(NAPPA)，其中每个球状蛋白都被筛选出与室颤患者的反应性 塞拉。我们的假设是，球虫蛋白质组的这次调查将发现新的球状相抗原。 对更多的患者有反应，特别是TP和/或CF血清阴性的危重患者 抗原。一旦我们确定了一小部分血清反应抗原，它们就可以被合并到现有的抗原中。 准备或单独的可靠、一致、准确的检测，将增加阳性诊断的数量 在患有早期疾病的急性病患者中。

PROJECT SUMMARY As of 2017, coccidioidomycosis (Cocci) also known as Valley Fever (VF) was designated a reportable disease in 22 states (1). The majority of the cases in the US occur in southern Arizona and California where the population is increasing with influx of naïve individuals, in particular, elderly retirees. Since Coccidioides sp, can cause significant morbidity and mortality in fully immunocompetent hosts, this represents an important threat to these regions where >10% percent of the US population resides, not including substantial tourist or part-time resident populations in Arizona and California. Importantly, the reporting criteria require a positive laboratory test for inclusion, yet ~50% of patients–who will eventually test positive–test negative while acutely ill (2). Testing has low sensitivity and specificity leaving sick patients and their health care providers without an accurate diagnosis. Thus, there is a profound need for better diagnostic approaches to properly identify Cocci patients to ensure appropriate follow-up and therapy ensues. Coccidioides spp. grow as mycelia in the desert soils and produce spores (arthroconidia) to survive during times of adverse growth conditions. When this soil is disturbed, arthroconidia aerosolize and are inhaled to initiate infection. Inside the host, the spores transform into spherules containing endospores which grow in the lung. When spherules burst, endospores, are released and can each disseminate to form a new spherule. Clinical diagnosis is difficult because patients’ symptoms resemble other bacterial and viral pneumonias. Laboratory diagnosis often relies solely on serology, but ~50% of patients do not test positive while they are acutely ill which results in inaccurate diagnosis an inappropriate treatment. We propose that it is time for a re- evaluation of seroreactive coccidioidal antigens. For the past six decades, serologic diagnosis of coccidioidomycosis has relied on IgM responses to tube precipitin (TP) and IgG responses to complement fixation (CF) antigens. Since these antigens are primarily expressed in mycelia, not in spherules which is the fungal form that grows in the host, it is not surprising that many patients are seronegative because they have not yet generated antibodies to spherule antigens during acute pulmonary illness. The GOAL of this proposal is to identify new coccidioidal antigens that react with acutely infected patient sera. The objectives are to utilize the recently published Coccidioides spp. proteome to create a nucleic acid programmable protein array (NAPPA) in which every coccidioidal protein is screened for reactivity with VF patient sera. Our hypothesis is that this survey of the Coccidioides proteome will reveal new spherule-phase antigens that are reactive with more patients, especially acutely ill patients who are seronegative for TP and/or CF antigens. Once we identify a small panel of seroreactive antigens, they can be incorporated into existing antigen preparations or a separate reliable, consistent, accurate test that will increase the number of positive diagnoses in acutely ill patients with early disease.