Multiplexed quantification of circulating peptidomic signatures for EBOLA early diagnosis
用于埃博拉早期诊断的循环肽组特征的多重定量
基本信息
- 批准号:9387209
- 负责人:
- 金额:$ 17.41万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2017
- 资助国家:美国
- 起止时间:2017-07-07 至 2019-06-30
- 项目状态:已结题
- 来源:
- 关键词:AddressAfricaAmplifiersAnimalsAntibodiesAreaBiological AssayBiological MarkersCalibrationCaringCategoriesCenters for Disease Control and Prevention (U.S.)CharacteristicsCleaved cellCohort StudiesCommunicable DiseasesContainmentCustomDataDemocratic Republic of the CongoDetectionDiagnosisDiagnosticDiagnostic testsDigestionDiseaseDisease OutbreaksEarly DiagnosisEbola Hemorrhagic FeverEbola virusEmergency SituationEndemic DiseasesEpidemicEquipmentEventExhibitsFiloviridaeFilovirusFormic AcidsFrankfurt-Marburg Syndrome VirusFutureHourHumanHuman ResourcesHydrolysisImmune responseIndividualInfectionInfectious AgentInternationalLaboratoriesLassa FeverMALDI-TOF Mass SpectrometryMalariaMass Spectrum AnalysisMediatingMedical ResearchMeningitisMethodologyMethodsModelingMolecularMorbidity - disease rateNanostructuresNanotechnologyPeptidesPhysiciansPilot ProjectsPlasmaPre-Clinical ModelPreparationPrimatesProceduresProteinsProtocols documentationRNARecombinant ProteinsRecombinantsRecoveryReproducibilityResearch InstituteResolutionRestonSafetySamplingSensitivity and SpecificitySerumSerum ProteinsSignal TransductionSiliconSiteSpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationSudanSurfaceSymptomsSystemTestingTimeTyphoid FeverUnited States Food and Drug AdministrationUnited States National Institutes of HealthVariantViralViral AntigensViral Hemorrhagic FeversViral load measurementVirionVirus Diseasesantibody conjugatebaseclinical applicationdiagnostic assaydisease diagnosiseffective therapyforesthospital laboratoriesimprovedinstrumentionizationmicrowave electromagnetic radiationminiaturizemortalitymultiplex detectionnanodisknanostructurednew technologynonhuman primatenovelpathogenpoint of carepre-clinicalprotein complexrapid diagnosisstable isotopeviral RNAviral detection
项目摘要
ABSTRACT
Ebola virus (EBOV) is classified as a filamentous, enveloped, non-segmented negative-sense RNA carrying
Category A Priority Pathogen by the NIH and CDC. Infections with EBOV cause severe hemorrhagic fever and
high mortality in humans and non-human primates (NHPs). During the Ebola outbreak of late 2014 in West Africa,
the lack of laboratory tests for early Ebola-virus-disease (EVD) detection (preferably before the onset of
symptoms) and lack of effective treatments limited the containment and management of the eventual epidemic.
To address this critical need for early EVD diagnosis, we have recently customized a platform we developed to
enhance biomarker detection to allow sensitive and specific EBOV biomarker quantification in serum samples.
Our method uses antibody-modified porous silicon nanodisks (pSiNDs) and high-throughput, high-resolution
mass spectrometry (MS) – together referred to as pSiND-MS – to greatly enhance the specificity and sensitivity
of target peptide detection from complex protein samples. In our customized EVD assay, these pSiNDs are
functionalized with custom antibodies to specific peptides of the EBOV matrix protein VP40 in order to efficiently
capture these peptides from non-enzymatically-digested serum samples of EBOV-infected hosts. Notably, the
starting non-enzymatic digestion step of this assay is used to both inactivate live EBOV particles and to site-
specifically digest all serum proteins for subsequent pSiND-MS analysis. We have observed that VP40 peptides
are increased in the plasma of infected animals even before EBOV RNA is detectable with currently available
diagnostic assays. We have identified EBOV- and EBOV-species-specific VP40 peptides, and hypothesize that
multiplex detection of our two selected VP40 peptides will robustly indicate EVD, EBOV species and EBOV load
in potentially-infected individuals. We will thus apply pSiND-MS to quantify low-abundance VP40 peptides for
early diagnosis of EBOV using a rapid, single-test assay, where we propose to: 1) optimize the pSiND-MS
platform for detection of these biomarkers and 2) apply pSiND-MS to longitudinally quantify VP40 levels in sera
from EBOV-infected non-human primates at the U.S. Army Medical Research Institute of Infectious Diseases, to
produce an informative preclinical EBOV-detection profile.
Major advantages of our approach include the simplicity and safety of sample preparation; the use of matrix-
assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS equipment found in most hospitals and
laboratories (and future use of simplified, miniaturized MS systems); the potential to read the assay on MS/MS
instruments for marker sequence confirmation; the specificity and sensitivity of biomarker detection; the use of
rapidly-customizable, modular multiplex assay; and rapid diagnosis (2hrs from sample). These characteristics
are ideal for a point-of-need application. Successful completion of this proposal will provide a novel technology
that can overcome significant hurdles of current diagnostic methodologies for Ebola and other infectious agents,
potentially transforming the protocols for care, treatment, and containment of future epidemics.
摘要
埃博拉病毒(Ebola Virus,EBOV)是一种丝状、囊膜、非节段性负义RNA载体
美国国立卫生研究院和美国疾病控制与预防中心的A类优先病原体。感染EBOV会导致严重的出血热和
人类和非人类灵长类动物(NHP)的高死亡率。在2014年底西非爆发埃博拉疫情期间,
缺乏早期埃博拉病毒病(EVD)检测的实验室检测(最好是在
病症)和缺乏有效的治疗方法限制了对最终疫情的控制和管理。
为了满足对早期EVD诊断的这一迫切需求,我们最近定制了一个我们开发的平台
增强生物标记物检测,以便在血清样本中进行敏感和特异的EBOV生物标记物定量。
我们的方法使用抗体修饰的多孔硅纳米盘(PSiND)和高通量、高分辨率
质谱学(MS)-统称为pSiND-MS-极大地提高了特异性和灵敏度
从复杂蛋白质样品中检测目标多肽的方法。在我们定制的EVD检测中,这些pSiND是
使用针对EBOV基质蛋白VP40的特定多肽的定制抗体来功能化,以便有效地
从EBOV感染宿主的非酶消化血清样本中捕获这些多肽。值得注意的是,
本试验的非酶消化步骤用于灭活活的EBOV颗粒和定位EBOV颗粒。
为后续的pSiND-MS分析专门消化所有血清蛋白。我们观察到VP40多肽
甚至在目前可用的EBOV RNA检测到之前,感染动物的血浆中的EBOV就增加了
诊断化验。我们已经确定了EBOV和EBOV物种特异性的VP40多肽,并假设
我们选择的两个VP40多肽的多重检测将有力地指示EVD、EBOV种类和EBOV载量
在可能感染的个体中。因此,我们将应用pSiND-MS来量化低丰度的VP40多肽
使用快速、单次检测方法进行EBOV早期诊断,我们建议:1)优化pSiND-MS
这些生物标志物的检测平台和2)应用pSiND-MS纵向定量血清中的VP40水平
从美国陆军传染病医学研究所感染EBOV的非人类灵长类动物,到
制作一份信息丰富的临床前EBOV检测档案。
我们方法的主要优点包括样品制备的简单性和安全性;使用基质-
辅助激光解吸/电离飞行时间(MALDI-TOF)MS设备在大多数医院和
实验室(以及简化、小型化MS系统的未来使用);在MS/MS上读取化验结果的可能性
标记序列确认的仪器;生物标记检测的特异性和敏感性;
可快速定制的模块化多重分析;以及快速诊断(从样本开始2小时)。这些特点
是按需应用程序的理想选择。这项提案的成功完成将提供一项新的技术
这可以克服目前埃博拉和其他感染性病原体诊断方法的重大障碍,
有可能改变护理、治疗和遏制未来流行病的方案。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Tony Y. Hu其他文献
IP-MS Analysis of ESX-5 and ESX-1 Substrates Enables Mycobacterial Species Identification
ESX-5 和 ESX-1 底物的 IP-MS 分析可实现分枝杆菌菌种鉴定
- DOI:
- 发表时间:
2020 - 期刊:
- 影响因子:0
- 作者:
Qingbo Shu;Meena U Rajagopal;Jia Fan;Lingpeng Zhan;Xiangxing Kong;Yifan He;Suwatchareeporn Rotcheewaphan;Christopher J. Lyon;W. Sha;A. Zelazny;Tony Y. Hu - 通讯作者:
Tony Y. Hu
Phenotypic plasticity and secretory heterogeneity in subpopulations derived from single cancer cell
源自单个癌细胞的亚群中的表型可塑性和分泌异质性
- DOI:
10.1016/j.apsb.2025.02.039 - 发表时间:
2025-05-01 - 期刊:
- 影响因子:14.600
- 作者:
Zhun Lin;Siping Liang;Zhe Pu;Zhengyu Zou;Luxuan He;Christopher J. Lyon;Yuanqing Zhang;Tony Y. Hu;Minhao Wu - 通讯作者:
Minhao Wu
Blood-Based microRNA Biomarker Signature of Early-Stage Pancreatic Ductal Adenocarcinoma With Lead-Time Trajectory in Prediagnostic Samples
- DOI:
10.1016/j.gastha.2024.08.002 - 发表时间:
2024-01-01 - 期刊:
- 影响因子:
- 作者:
Warapen Treekitkarnmongkol;Jianliang Dai;Suyu Liu;Deivendran Sankaran;Tristian Nguyen;Seetharaman Balasenthil;Mark W. Hurd;Meng Chen;Hiroshi Katayama;Sinchita Roy-Chowdhuri;George A. Calin;Randall E. Brand;Paul D. Lampe;Tony Y. Hu;Anirban Maitra;Eugene J. Koay;Ann M. Killary;Subrata Sen - 通讯作者:
Subrata Sen
Recent advances in the bench-to-bedside translation of cancer nanomedicines
癌症纳米医学从实验室到临床转化的最新进展
- DOI:
10.1016/j.apsb.2024.12.007 - 发表时间:
2025-01-01 - 期刊:
- 影响因子:14.600
- 作者:
Yang Liu;Yinchao Zhang;Huikai Li;Tony Y. Hu - 通讯作者:
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Decoding the blood peptidome as a new biomarker resource for cancer detection
解码血液肽组作为癌症检测的新生物标志物资源
- DOI:
10.15406/mojpb.2016.03.00099 - 发表时间:
2016 - 期刊:
- 影响因子:0
- 作者:
Yaojun Li;Tony Y. Hu - 通讯作者:
Tony Y. Hu
Tony Y. Hu的其他文献
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{{ truncateString('Tony Y. Hu', 18)}}的其他基金
Multiplexed detection of cell-free M. Tuberculosis DNA and its drug-resistant variants in blood
血液中无细胞结核分枝杆菌 DNA 及其耐药变异体的多重检测
- 批准号:
10639855 - 财政年份:2023
- 资助金额:
$ 17.41万 - 项目类别:
Quantification of brain-derived extracellular vesicle microRNAs in blood by a liposome-mediated CRISPR assay for traumatic brain injury detection
通过脂质体介导的 CRISPR 测定对血液中脑源性细胞外囊泡 microRNA 进行定量,用于检测创伤性脑损伤
- 批准号:
10575436 - 财政年份:2022
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A nanopore biosensor for leveling Mtb antigens in blood
用于平衡血液中 Mtb 抗原的纳米孔生物传感器
- 批准号:
10646134 - 财政年份:2022
- 资助金额:
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Digital Nanoplasmonic Quantification of Tumor-derived Extracellular Vesicles in Plasma Microsamples
血浆微样品中肿瘤源性细胞外囊泡的数字纳米等离子体定量
- 批准号:
10684737 - 财政年份:2020
- 资助金额:
$ 17.41万 - 项目类别:
Digital Nanoplasmonic Quantification of Tumor-derived Extracellular Vesicles in Plasma Microsamples
血浆微样品中肿瘤源性细胞外囊泡的数字纳米等离子体定量
- 批准号:
10461970 - 财政年份:2020
- 资助金额:
$ 17.41万 - 项目类别:
Digital Nanoplasmonic Quantification of Tumor-derived Extracellular Vesicles in Plasma Microsamples
血浆微样品中肿瘤源性细胞外囊泡的数字纳米等离子体定量
- 批准号:
10269902 - 财政年份:2020
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$ 17.41万 - 项目类别:
Detecting pathogen and host factors on extracellular vesicles for pediatric TB diagnosis and management
检测细胞外囊泡上的病原体和宿主因子,用于儿童结核病的诊断和管理
- 批准号:
10753281 - 财政年份:2017
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$ 17.41万 - 项目类别:
Direct quantitation of the circulating Mtb-peptidome for pediatric TB management
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- 批准号:
9333558 - 财政年份:2017
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- 批准号:
9241942 - 财政年份:2016
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