Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, 20110, USA.
Queensland University of Technology, Brisbane, Australia.
Sci Rep. 2020 Nov 9;10(1):19340. doi: 10.1038/s41598-020-75051-3.
Mass spectrometry enhanced by nanotechnology can achieve previously unattainable sensitivity for characterizing urinary pathogen-derived peptides. We utilized mass spectrometry enhanced by affinity hydrogel particles (analytical sensitivity = 2.5 pg/mL) to study tick pathogen-specific proteins shed in the urine of patients with (1) erythema migrans rash and acute symptoms, (2) post treatment Lyme disease syndrome (PTLDS), and (3) clinical suspicion of tick-borne illnesses (TBI). Targeted pathogens were Borrelia, Babesia, Anaplasma, Rickettsia, Ehrlichia, Bartonella, Francisella, Powassan virus, tick-borne encephalitis virus, and Colorado tick fever virus. Specificity was defined by 100% amino acid sequence identity with tick-borne pathogen proteins, evolutionary taxonomic verification for related pathogens, and no identity with human or other organisms. Using a cut off of two pathogen peptides, 9/10 acute Lyme Borreliosis patients resulted positive, while we identified zero false positive in 250 controls. Two or more pathogen peptides were identified in 40% of samples from PTLDS and TBI patients (categories 2 and 3 above, n = 59/148). Collectively, 279 distinct unique tick-borne pathogen derived peptides were identified. The number of pathogen specific peptides was directly correlated with presence or absence of symptoms reported by patients (ordinal regression pseudo-R = 0.392, p = 0.010). Enhanced mass spectrometry is a new tool for studying tick-borne pathogen infections.
纳米技术增强的质谱分析可实现对尿源性病原体衍生肽进行特征分析的前所未有的灵敏度。我们利用亲和水凝胶颗粒增强的质谱分析（分析灵敏度为 2.5 pg/mL）来研究在患有（1）游走性红斑皮疹和急性症状、（2）莱姆病后综合征（PTLDS）和（3）疑似 tick-borne 疾病（TBI）的患者尿液中释放的 tick 病原体特异性蛋白。靶向病原体为 Borrelia、Babesia、Anaplasma、Rickettsia、Ehrlichia、Bartonella、Francisella、Powassan 病毒、蜱传脑炎病毒和科罗拉多蜱热病毒。特异性通过与 tick-borne 病原体蛋白的 100%氨基酸序列同一性、相关病原体的进化分类学验证以及与人类或其他生物体无同一性来定义。使用两个病原体肽的截止值，10/10 例急性莱姆 Borreliosis 患者呈阳性，而在 250 例对照中我们未发现假阳性。PTLDS 和 TBI 患者（上述类别 2 和 3，n = 59/148）40%的样本中鉴定出两个或更多病原体肽。总共鉴定出 279 个独特的 tick-borne 病原体衍生肽。病原体特异性肽的数量与患者报告的症状的存在或不存在直接相关（有序回归伪 R = 0.392，p = 0.010）。增强的质谱分析是研究 tick-borne 病原体感染的新工具。
