Nebbak A, El Hamzaoui B, Berenger J-M, Bitam I, Raoult D, Almeras L, Parola P
Aix Marseille Université, Unité de Recherche en Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63, CNRS 7278, IRD 198 (Dakar, Sénégal), Inserm 1095, Institut Hospitalo-Universitaire Méditerranée Infection, Marseille, France.
Laboratoire de Biodiversité et Environnement: Interactions génomes, Faculté des Sciences Biologiques, Université des Sciences et de la Technologie Houari Boumediene (USTHB), Bab Ezzouar, Algeria.
Med Vet Entomol. 2017 Dec;31(4):438-448. doi: 10.1111/mve.12250. Epub 2017 Jul 19.
Ticks and fleas are vectors for numerous human and animal pathogens. Controlling them, which is important in combating such diseases, requires accurate identification, to distinguish between vector and non-vector species. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was applied to the rapid identification of arthropods. The growth of this promising tool, however, requires guidelines to be established. To this end, standardization protocols were applied to species of Rhipicephalus sanguineus (Ixodida: Ixodidae) Latreille and Ctenocephalides felis felis (Siphonaptera: Pulicidae) Bouché, including the automation of sample homogenization using two homogenizer devices, and varied sample preservation modes for a period of 1-6 months. The MS spectra were then compared with those obtained from manual pestle grinding, the standard homogenization method. Both automated methods generated intense, reproducible MS spectra from fresh specimens. Frozen storage methods appeared to represent the best preservation mode, for up to 6 months, while storage in ethanol is also possible, with some caveats for tick specimens. Carnoy's buffer, however, was shown to be less compatible with MS analysis for the purpose of identifying ticks or fleas. These standard protocols for MALDI-TOF MS arthropod identification should be complemented by additional MS spectrum quality controls, to generalize their use in monitoring arthropods of medical interest.
蜱虫和跳蚤是多种人类和动物病原体的传播媒介。对其进行控制在对抗此类疾病方面至关重要，而这需要准确识别，以区分传播媒介物种和非传播媒介物种。最近，基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）被应用于节肢动物的快速识别。然而，这种有前景的工具的发展需要建立相关指南。为此，标准化方案应用于血红扇头蜱（硬蜱目：硬蜱科）拉特雷尔种和猫栉首蚤指名亚种（蚤目：蚤科）布歇种，包括使用两种匀浆器实现样本匀浆自动化，以及1至6个月的不同样本保存模式。然后将质谱图与通过标准匀浆方法——手动杵磨获得的质谱图进行比较。两种自动化方法均能从新鲜标本中生成强烈且可重复的质谱图。冷冻保存方法似乎是长达6个月的最佳保存模式，而乙醇保存也可行，但蜱虫标本需注意一些事项。然而，事实证明，对于蜱虫或跳蚤的识别而言，卡诺伊缓冲液与质谱分析的兼容性较差。这些用于MALDI-TOF MS节肢动物识别的标准方案应辅以额外的质谱图质量控制，以便将其应用推广至监测具有医学意义的节肢动物。
