Cantero Paola, Ehret-Sabatier Laurence, Lenormand Cédric, Hansmann Yves, Sauleau Erik, Zilliox Laurence, Westermann Benoit, Jaulhac Benoit, Mutter Didier, Barthel Cathy, Perdu-Alloy Pauline, Martinot Martin, Lipsker Dan, Boulanger Nathalie
Laboratoire de Spectrométrie de Masse BioOrganique, Université de Strasbourg, Centre National de la Recherche Scientifique, Institut Pluridisciplinaire Hubert Curien, Unité Mixte de Recherche, Strasbourg, France; Infrastructure Nationale de Protéomique ProFI, Strasbourg, France.
Faculté de Médecine, Université de Strasbourg et Clinique Dermatologique, Hôpital Universitaire de Strasbourg, Strasbourg, France; UR3073-Pathogen-Host- Arthropod Vectors Interactions-Group Borrelia, Fédération de Médecine Translationnelle, Université de Strasbourg, Strasbourg, France.
Clin Microbiol Infect. 2025 Jan;31(1):78-86. doi: 10.1016/j.cmi.2024.10.014. Epub 2024 Oct 23.
We have developed targeted proteomics in the context of Lyme borreliosis (LM) as a new direct diagnostic tool for detecting Borrelia proteins in the skin of patients with erythema migrans. If satisfactory, this proteomic technique could be used in addition to culture and/or PCR for disseminated infections where Borrelia detection is essential to demonstrate active infection. In these infections, the diagnosis is indirect and relies mainly on serology.
We recruited 46 patients with LM and 11 controls and collected two skin biopsies from each patient. One biopsy was used for Borrelia burgdorferi sensu lato PCR and culture and the other one was for targeted mass-spectrometry-based proteomics. Six markers of infection were selected for proteomics: Outer surface protein C (OspC), flagellin, enolase, lipoprotein gi|365823350, decorin binding protein A, and glyceraldehyde-3-phosphate dehydrogenase.
Culturing Borrelia from the biopsies increased the sensitivity of the methods. Among the patients included for analysis, 61% (28 patients), 61% (28), and 46% (21) were detected as positive by proteomics, PCR, and culture, respectively. PCR and proteomics were complementary. OspC and flagellin were the most frequently detected protein markers of infection by proteomics, which in some patients, detected up to nine peptides for the flagellin.
It is possible to identify bacterial makers from the skin by proteomics. Our approach can be used to diagnose tick-borne diseases such as LM.
clinicaltrials.gov identifier: NCT02414789.
我们已在莱姆病（LM）背景下开发了靶向蛋白质组学，作为一种新的直接诊断工具，用于检测游走性红斑患者皮肤中的疏螺旋体蛋白。如果结果令人满意，这种蛋白质组学技术可用于除培养和/或聚合酶链反应（PCR）之外的播散性感染诊断，在这些感染中，疏螺旋体检测对于证明存在活动性感染至关重要。在这些感染中，诊断是间接的，主要依靠血清学。
我们招募了46例LM患者和11名对照，并从每位患者身上采集了两份皮肤活检样本。一份活检样本用于伯氏疏螺旋体广义PCR检测和培养，另一份用于基于靶向质谱的蛋白质组学分析。为蛋白质组学选择了六个感染标志物：外表面蛋白C（OspC）、鞭毛蛋白、烯醇化酶、脂蛋白gi|365823350、饰胶蛋白聚糖结合蛋白A和甘油醛-3-磷酸脱氢酶。
从活检样本中培养疏螺旋体提高了检测方法的灵敏度。在纳入分析的患者中，蛋白质组学、PCR和培养检测出的阳性率分别为61%（28例患者）、61%（28例）和46%（21例）。PCR和蛋白质组学具有互补性。OspC和鞭毛蛋白是蛋白质组学检测中最常发现的感染蛋白标志物，在一些患者中，鞭毛蛋白检测到多达九个肽段。
通过蛋白质组学从皮肤中鉴定细菌标志物是可行的。我们的方法可用于诊断蜱传疾病，如LM。
clinicaltrials.gov标识符：NCT02414789。
