Boyer Pierre H, Boulanger Nathalie, Nebbak Amira, Collin Elodie, Jaulhac Benoit, Almeras Lionel
Early Bacterial Virulence: Lyme borreliosis Group, Université de Strasbourg, Fédération de Médecine Translationnelle de Strasbourg, VBP EA 7290, Strasbourg, France.
French National Reference Center for Borrelia, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.
PLoS One. 2017 Sep 26;12(9):e0185430. doi: 10.1371/journal.pone.0185430. eCollection 2017.
Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been demonstrated to be useful for tick identification at the species level. More recently, this tool has been successfully applied for the detection of bacterial pathogens directly in tick vectors. The present work has assessed the detection of Borrelia burgdorferi sensu lato in Ixodes ricinus tick vector by MALDI-TOF MS. To this aim, experimental infection model of I. ricinus ticks by B. afzelii was carried out and specimens collected in the field were also included in the study. Borrelia infectious status of I. ricinus ticks was molecularly controlled using half-idiosome to classify specimens. Among the 39 ticks engorged on infected mice, 14 were confirmed to be infected by B. afzelii. For field collection, 14.8% (n = 12/81) I. ricinus ticks were validated molecularly as infected by B. burgdorferi sl. To determine the body part allowing the detection of MS protein profile changes between non-infected and B. afzelii infected specimens, ticks were dissected in three compartments (i.e. 4 legs, capitulum and half-idiosome) prior to MS analysis. Highly reproducible MS spectra were obtained for I. ricinus ticks according to the compartment tested and their infectious status. However, no MS profile change was found when paired body part comparison between non-infected and B. afzelii infected specimens was made. Statistical analyses did not succeed to discover, per body part, specific MS peaks distinguishing Borrelia-infected from non-infected ticks whatever their origins, laboratory reared or field collected. Despite the unsuccessful of MALDI-TOF MS to classify tick specimens according to their B. afzelii infectious status, this proteomic tool remains a promising method for rapid, economic and accurate identification of tick species. Moreover, the singularity of MS spectra between legs and half-idiosome of I. ricinus could be used to reinforce this proteomic identification by submission of both these compartments to MS.
基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)已被证明可用于蜱虫种类水平的鉴定。最近,该工具已成功应用于直接在蜱虫载体中检测细菌病原体。本研究评估了通过MALDI-TOF MS检测蓖麻硬蜱载体中广义伯氏疏螺旋体。为此,进行了阿氏疏螺旋体对蓖麻硬蜱的实验感染模型,并且研究中还纳入了野外采集的标本。使用半体对蓖麻硬蜱的伯氏疏螺旋体感染状况进行分子控制以对标本进行分类。在39只叮咬感染小鼠的蜱虫中,有14只被证实感染了阿氏疏螺旋体。对于野外采集的标本,14.8%(n = 12/81)的蓖麻硬蜱在分子水平上被验证感染了广义伯氏疏螺旋体。为了确定能够检测未感染和阿氏疏螺旋体感染标本之间质谱蛋白质谱变化的身体部位,在进行质谱分析之前,将蜱虫解剖为三个部分(即4条腿、头部和半体)。根据所测试的部分及其感染状况,获得了蓖麻硬蜱高度可重复的质谱图。然而,在未感染和阿氏疏螺旋体感染标本的配对身体部位比较中未发现质谱图变化。统计分析未能按身体部位发现区分感染伯氏疏螺旋体和未感染蜱虫的特定质谱峰,无论其来源是实验室饲养还是野外采集。尽管MALDI-TOF MS未能根据蜱虫的阿氏疏螺旋体感染状况对蜱虫标本进行分类,但这种蛋白质组学工具仍然是一种用于快速、经济和准确鉴定蜱虫种类的有前景的方法。此外,蓖麻硬蜱腿部和半体之间质谱图的独特性可用于通过将这两个部分都进行质谱分析来加强这种蛋白质组学鉴定。