A Cost-Effective High-Speed Clinical Diagnostics Instrument for Large Population Screening Based on Novel Liquid AP-MALDI MS Technology

基于EPSRC先前资助的研究（EP/L006227/1）“开发一种新型MALDI质谱仪和高灵敏度产生多电荷离子的技术”，以及随后对该新技术的初步开发，该项目将开发一种新的仪器，专门满足现代医学对临床诊断的关键要求，特别是在COVID-19等新流行病时代。临床样品中微生物的准确和快速表征对于启动最佳治疗和限制大流行的爆发至关重要。准确性和时间都是患者获得最佳治疗结果的关键，最大限度地减少恢复时间，更重要的是减少发病率和死亡率。特别是，正确和快速地鉴定新发现的微生物病原体或耐药菌株对患者的康复至关重要。与大规模检测能力相结合，它还将使全球更好地应对（微生物）传染病。基质辅助激光解吸/电离（MALDI）质谱（MS）生物谱分析（生物分型）最近被确立为一种优于经典临床微生物学检测的方法，用于鉴定临床相关微生物，大大提高了分类准确性和分析速度。这已经导致了两个FDA批准的微生物检测和鉴定系统MALDI质谱生物分型。在提议的项目中，该方法将通过利用多种带电离子及其与脂质和其他生物分子在台式MS/MS仪器上的共同分析而得到实质性的进步，该仪器专门针对大规模，廉价的临床分析进行优化，从而导致下一代卓越的MALDI MS生物分型，用于临床使用和大规模测试。这种新仪器和相关技术的独特之处在于其高速度、高成本效益和高特异性的MS/MS测序。结合质谱法无与伦比的分辨率，这项新技术将是诊断测试的一个阶阶变化，它允许在同一次测试中测试多种疾病，并且对新疾病具有高度适应性，而无需开发针对疾病/微生物的测试试剂，难以采购，因此价格昂贵，特别是对于新发现的疾病（参见COVID - 19）。目标是在高检测精度和低样品成本的情况下达到每天100,000个样品的吞吐量水平。通过与生物制药行业和分析仪器制造商的合作，以及从bbsrc资助的研究中，我们发现我们的新型AP-MALDI质谱离子源为快速分析提供了一个敏感的平台，具有用于微生物疾病早期检测的潜力。拟议的项目将建立在这些初步数据的基础上，为未来的临床使用开发一种新的定制仪器，并探索（与目前的MALDI MS生物分型仪相比）在以下方面的优势(a)速度，(b)消除生物基质背景，(c)卓越的MS/MS分析，(d)更大的离子信号稳定性，(e)多路复用能力和(f)更简单（且具有成本效益）但更灵活的样品制备，这项新技术可以提供。

Based on previously funded EPSRC research (EP/L006227/1) for the 'Development of a novel MALDI mass spectrometer and technology for the generation of multiply charged ions at high sensitivity' and subsequent initial exploitation of this new technology, the proposed project will develop a new instrument that specifically fulfils key requirements in clinical diagnostics as demanded by modern medicine, in particular in the age of new pandemics such as COVID-19.Accurate and fast characterisation of microorganisms in clinical samples are crucial for initiating optimal treatment and limiting the outbreaks of pandemics. Both accuracy and time are key to the best treatment outcome for the patient, minimising the time to recovery and more importantly minimising morbidity and mortality. In particular, the correct and rapid identification of newly discovered microbial pathogens or antimicrobial-resistant strains is important for the patient's recovery. Combined with the capability of large-scale testing, it will also allow for a better global response to (microbial) infectious diseases.Matrix-assisted laser desorption/ionisation (MALDI) mass spectrometry (MS) profiling of organisms (biotyping) has recently been established as a superior method to classical clinical microbiology assays for the identification of clinically relevant microbes with substantially increased classification accuracy and speed of analysis. This has already led to two FDA approved systems for microbial detection and identification by MALDI MS biotyping. In the proposed project, this methodology will be substantially advanced by exploiting multiply charged ions and their co-analysis with lipids and other biomolecules on a bench-top MS/MS instrument specifically optimised for large-scale, inexpensive clinical analyses, thus leading to the next generation of superior MALDI MS biotyping for clinical use and mass testing.The unique features of this new instrument and the associated technology will be high speed, cost-effectiveness, and high specificity by MS/MS sequencing. Combined with the unrivalled resolution of mass spectrometry this new technology will be a step-change in diagnostic testing by allowing the testing of multiple diseases within the same test run as well as being highly adaptable to new diseases without the need to develop test reagents that are disease/microbe-specific, difficult to source and therefore expensive, in particular for newly discovered diseases (cf. COVID 19). The aim is to reach a throughput level of 100,000 samples per day at high detection accuracy and low cost per sample.From collaborating with the biopharmaceutical industry and analytical instrument manufacturers, and from research of a BBSRC-funded grant, we found that our novel AP-MALDI MS ion source provides a sensitive platform for rapid assay analysis with the potential for use in the early detection of microbial diseases. The proposed project will build on this preliminary data, develop a new tailor-made instrument for future clinical use and explore the advantages (compared to current MALDI MS biotypers) in (a) speed, (b) the elimination of biological matrix background, (c) superior MS/MS analysis, (d) greater ion signal stability, (e) multiplexing capability and (f) the simpler (and cost-effective) but more flexible sample preparation that this new technology can offer.