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首页> 外文期刊>Microbiology >Visualization of Borrelia burgdorferi sensu lato by fluorescence in situ hybridization (FISH) on whole-body sections of Ixodes ricinus ticks and gerbil skin biopsies
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Visualization of Borrelia burgdorferi sensu lato by fluorescence in situ hybridization (FISH) on whole-body sections of Ixodes ricinus ticks and gerbil skin biopsies

机译：通过荧光在Ixodes Ricinus蜱和沙鼠皮肤活检的全身杂交（鱼类）上荧光的荧光杂交（鱼）的可视化

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The objective of this study was to visualize borreliae directly in whole-body sections of Ixodes ricinus by fluorescence in situ hybridization (FISH). Borrelia afzelii mono-infected or Borrelia burgdorferi sensu stricto (ss)/B. afzelii double-infected nymphs were fixed, embedded in cold polymerizing resin and sectioned. The same sample processing was applied to skin biopsies taken from a Mongolian gerbil after an infectious tick-bite. FISH was carried out using 16S-rRNA-directed, fluorescence-labelled oligonucleotide probes specific for the genus Borrelia and specific within the group of Lyme borreliosis-associated genospecies B. afzelii, B. burgdorferi ss, Borrelia garinii and Borrelia valaisiana. Sensitivity and specificity of the newly designed probes were evaluated using PCR, dot-blot hybridizations and FISH. Despite significant autofluorescence of certain tick tissues (which allowed good histological orientation within the sections), borreliae showing the typical spirochaetal morphotype were clearly visible in five out of six putatively infected ticks. These findings were confirmed by electron microscopy of ticks from the same infected batch as used for FISH. Attempts to produce ticks infected by two different Borrelia genospecies were not successful. FISH on whole-body sections of resin-embedded ticks offers the possibility of visualizing and identifying borreliae within tick tissues. This technique has great potential for the investigation of the transmission of bacteria or other micro-organisms by arthropod vectors. Furthermore, clear visualization of single spirochaetes distributed along subcutaneous fat cell membranes in gerbil skin biopsies suggests that FISH might also be suitable for the detection of borreliae in clinical tissue specimens.
机译：本研究的目的是通过原位杂交（鱼类）直接在Ixodes Ricinus的全身部分中可视化伯雷雷氏症。 Borrelia Afzelii单声道或博福雷莱伯戈多弗Sensuto（SS）/ b。将Afzelii双感染的若虫固定，嵌入冷聚合树脂和切片中。在感染性蜱咬咬合后，将相同的样品加工应用于从蒙古菌中取出的皮肤活组织检查。使用16s-rRNA定向的荧光标记的荧光标记的寡核苷酸探针进行鱼类，特异于博尔塞尔属，并且在莱利氏肾素病患者组中的特异性，B.Fzelii，B.Burgdorferi SS，Borrelia Garinii和Borrelia Valaisiana。使用PCR，点印迹杂交和鱼评估新设计探针的敏感性和特异性。尽管某些蜱组织的显着自发荧光（其允许在部分内的良好的组织学取向），但博洛雷莱显示出典型的螺旋形式Mor蝶型在六个令人沮丧的蜱中清晰可见。通过用于鱼的相同受感染的批料的电子显微镜通过电子显微镜证实了这些发现。试图产生被两次不同的博福雷亚生殖器生殖器感染的蜱没有成功。树脂嵌入式蜱虫的全身部分的鱼类提供了可视化和识别蜱组织内的博尔塞雷的可能性。该技术具有对节肢动物载体传播细菌或其他微生物的调查潜力。此外，沿着红霉皮肤活检沿皮下脂肪细胞膜分布的单螺螺旋体的清晰可视化表明鱼类可能也适用于临床组织标本中的博尔塞雷。

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