Hybridoma Laboratory, School of Biosciences, University of Exeter, Exeter, United Kingdom.
Adv Appl Microbiol. 2010;70:187-216. doi: 10.1016/S0065-2164(10)70006-X. Epub 2010 Mar 6.
Invasive aspergillosis (IA) caused by the fungus Aspergillus fumigatus is a frequent and life-threatening complication of chemotherapy and bone marrow transplantation with high rates of mortality and morbidity. Diagnosis of IA is complex and can only be confirmed by identification of the fungus in biopsy samples. Capturing tissue for diagnosis is in itself hazardous, and because of this many patients receive empirical antifungal treatment rather than undergo biopsy. However, the treatment carries with it significant side effects and is prohibitively expensive. Because of this, attempts have been made to develop specific and sensitive diagnostic tests that can be used to track the early onset of infection and permit rational administration of antifungal drugs. Early attempts at nonculture-based diagnosis using human immune serum to detect circulating Aspergillus antigens proved unreliable, and so focus turned to hybridoma technology and the use of monoclonal antibodies (MAbs) to detect signature molecules of infection. Detection of one such signature molecule, galactomannan (and associated galactomannoprotein molecules), forms the basis of the commercial Platelia enzyme immunoassay (EIA), an assay that has found widespread use in IA diagnosis. Nevertheless, concerns surrounding its accuracy mean that alternative strategies to diagnosis have been sought including detection of the fungal cell wall component (1-->3)-beta-d-glucan and polymerase chain reaction (PCR). The poor specificity of "panfungal" (1-->3)-beta-d-glucan tests and current lack of standardization of PCR assays have led to the recent development of next-generation MAb-based assays that detect surrogate markers of infection and that have been incorporated into "point-of-care" diagnostic devices. This chapter examines the development of antibody-antigen, (1-->3)-beta-d-glucan, and nucleic acid-based approaches to IA detection, current concerns surrounding accurate disease diagnosis, and how animal models of infection can be used to inform assay development and validation.
由烟曲霉引起的侵袭性曲霉病（IA）是化疗和骨髓移植的常见且危及生命的并发症，具有高死亡率和发病率。IA 的诊断很复杂，只能通过在活检样本中识别真菌来确认。采集用于诊断的组织本身就有危险，因此许多患者接受经验性抗真菌治疗，而不是进行活检。然而，这种治疗会带来严重的副作用，而且非常昂贵。因此，人们试图开发特异性和敏感性诊断测试，以便跟踪感染的早期发作，并允许合理使用抗真菌药物。早期使用人类免疫血清检测循环曲霉属抗原的非培养诊断尝试结果不可靠，因此研究重点转向杂交瘤技术和使用单克隆抗体（MAb）检测感染的特征分子。检测其中一种特征分子，半乳甘露聚糖（和相关的半乳甘露糖蛋白分子），构成了商业 Platelia 酶免疫分析（EIA）的基础，该分析已在 IA 诊断中得到广泛应用。然而，对其准确性的关注意味着已经寻求替代诊断策略，包括检测真菌细胞壁成分（1-->3）-β-d-葡聚糖和聚合酶链反应（PCR）。“泛真菌”（1-->3）-β-d-葡聚糖测试的特异性差和当前 PCR 测试缺乏标准化导致了新一代基于 MAb 的测试的发展，这些测试检测感染的替代标志物，并已纳入“即时”诊断设备。本章探讨了针对 IA 检测的抗体-抗原、（1-->3）-β-d-葡聚糖和核酸方法的发展、围绕准确疾病诊断的当前关注点，以及感染动物模型如何用于告知测试开发和验证。
