Using silicon-based biosensors to detect host protein signatures capable of differentiating between bacterial and viral infection at point-of-care

使用硅基生物传感器检测宿主蛋白质特征，能够在护理点区分细菌和病毒感染

• 批准号: 2621331
• 金额: --
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2621331
• 关键词: Using; silicon; based; biosensors; detect

The World Health Organisation has predicted that the rise of antimicrobial resistance (AMR) will claim up to 10 million lives by the year 2050[1]. One of the key factors driving AMR emergence is the lack of a rapid diagnostic device capable of differentiating between bacterial and viral infection. A bacterial culture test is the current "gold standard", however it presents with various draw backs such as long time-to-positive results and risk of sample contamination[2]. This leads to clinicians either delaying antibiotic treatment as they wait for confirmation or overprescribing broad spectrum antibiotics in cases of severe infection. It is then unsurprising to learn that around 50% of antibiotics are inappropriately prescribed[3]. The ideal diagnostic device would be rapid and available at point-of-care with high sensitivity and specificity, however challenges with current diagnostics make these characteristics difficult to achieve. Primarily, they mostly rely on optical-based methods of detection which usually require bulky and expensive laboratory equipment and so often makes these assays unavailable at point-of-care[4]. Additionally, direct detection of the causative pathogen may not be possible as some pathogens are incredibly invasive or inaccessible, especially in cases of tuberculosis. Finally, confirmation of a bacterial infection does not rule out a causative viral infection and vice versa which is a very common dynamic in upper respiratory infections[5].Host protein signatures could bypass all these challenges and various host protein signatures have already been cited in literature. Oved et al. (2015) discovered a 3 host protein signature composed of C-reactive protein (CRP), TNF-related apoptosis-inducing ligand (TRAIL) and Interferon gamma-induced protein-10 (IP-10) which is able to accurately differentiate between bacterial and viral infection[6]. However, it is estimated that over 90% of discovered host biomarkers have not yet been applied successfully at point-of-care[7]. Combining host protein signatures with already leveraged complementary metal-oxide semiconductor technology (CMOS) (the leading technology used to manufacture everyday electronics) would make such devices incredibly cheap and scalable[8]. CMOS technology is also compatible with ion sensitive field effect transistors (ISFETs), which can be used to detect changes in surface charge due to antigen-antibody binding and hence detect and quantify host proteins in real time.Research questions:-What specific host protein signature is able to differentiate between bacterial and viral infection with high sensitivity and specificity? -How does the sensitivity and specificity of the immunoassay immobilised on-chip compare to the "gold standard" protein detection technique?-What techniques are required to immobilise primary antibodies onto an array of ISFET sensors? -What is the on-chip limit of detection for that specific protein signature?Aims:-This project aims to develop an immunoassay specific to a host protein signature capable of differentiation between bacterial and viral infection. -The specific immunoassay will then be translated onto a silicon-based biosensor and the on-chip limit of detection will be established. -Finally, the assay will be validated against clinical samples and used to provide quantitative data around the infection state of the patient. References [1] M. E. A. de Kraker, A. J. Stewardson, and S. Harbarth, "Will 10 Million People Die a Year due to Antimicrobial Resistance by 2050?," PLOS Medicine, vol. 13, no. 11, p. e1002184, Nov. 2016, doi: 10.1371/JOURNAL.PMED.1002184.[2] J. C. Craig et al., "The accuracy of clinical symptoms and signs for the diagnosis of serious bacterial infection in young febrile children: prospective cohort study of 15 781 febrile illnesses," BMJ (Clinical research ed.), vol. 340, no. 7754, p. 1015, May 2010, doi: 10.1136/BMJ.C1594.[3] C. Giuliano, C. R. Patel, and

世界卫生组织预测，到2050年，抗菌素耐药性（AMR）的上升将夺去多达1000万人的生命[1]。推动AMR出现的关键因素之一是缺乏能够区分细菌和病毒感染的快速诊断设备。细菌培养测试是目前的“金标准”，但它存在各种缺点，如阳性结果的时间长和样品污染的风险[2]。这导致临床医生在等待确认时延迟抗生素治疗，或者在严重感染的情况下过度使用广谱抗生素。因此，了解到大约50%的抗生素是不适当的处方也就不足为奇了[3]。理想的诊断设备将是快速的，并且在护理点可用，具有高灵敏度和特异性，然而，当前诊断的挑战使得这些特征难以实现。首先，它们主要依赖于基于光学的检测方法，该方法通常需要庞大且昂贵的实验室设备，因此通常使得这些测定在护理点不可用[4]。此外，直接检测致病病原体可能是不可能的，因为一些病原体是令人难以置信的侵入性或难以接近的，特别是在结核病的情况下。最后，细菌感染的确认并不排除致病性病毒感染，反之亦然，这是上呼吸道感染中非常常见的动态[5]。宿主蛋白质特征可以绕过所有这些挑战，文献中已经引用了各种宿主蛋白质特征。Oved等人（2015）发现了由C反应蛋白（CRP）、TNF相关凋亡诱导配体（TRAIL）和干扰素γ诱导蛋白-10（IP-10）组成的3种宿主蛋白特征，能够准确区分细菌和病毒感染[6]。然而，据估计，超过90%的已发现的宿主生物标志物尚未成功应用于护理点[7]。将宿主蛋白质特征与已经利用的互补金属氧化物半导体技术（CMOS）（用于制造日常电子产品的领先技术）相结合，将使此类设备变得非常便宜和可扩展[8]。CMOS技术还与离子敏感场效应晶体管（ISFET）兼容，后者可用于检测抗原抗体结合引起的表面电荷变化，从而真实的实时检测和定量宿主蛋白。研究问题：-什么样的宿主蛋白特征能够以高灵敏度和特异性区分细菌和病毒感染？- 与“金标准”蛋白质检测技术相比，芯片上固定的免疫分析的灵敏度和特异性如何？将一抗固定在ISFET传感器阵列上需要什么技术？- 该特定蛋白质特征的芯片检测限是多少？目的：-该项目旨在开发一种特异于宿主蛋白质特征的免疫测定法，能够区分细菌和病毒感染。- 然后将特定的免疫测定转化到硅基生物传感器上，并建立芯片上的检测限。- 最后，将针对临床样本验证该测定，并用于提供患者感染状态的定量数据。参考文献[1] M. E. A. de Kraker，A. J. Stewardson和S. Harbarth，“到2050年，每年会有1000万人死于抗菌素耐药性吗？”“PLOS Medicine，第13卷，第11期，第e1002184页，2016年11月，doi：10.1371/JOURNAL.PMED.1002184。[2]J. C.克雷格等人，“临床症状和体征诊断年轻发热儿童严重细菌感染的准确性：15781例发热性疾病的前瞻性队列研究”，BMJ（临床研究编辑），第340卷，第7754号，第1015页，2010年5月，doi：10.1136/BMJ.C1594。[3]C.朱利亚诺，C。R.帕特尔，