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光现场混合作为植入物相关感染的诊断工具:关于附加值的试点研究

Catharina Scheuermann-Poley¹, Alexandra Wiessner², Judith Kikhney²

¹From the Trauma & Orthopedic Surgery, Septic & Reconstructive Surgery, Research and Treatment Centre Septic Defect Wounds, Federal Armed Forces of Germany, Bundeswehr (Military) Academic Hospital, Berlin, Germany.

Plastic and reconstructive surgery. Global open

|June 26, 2023

概括

光在位杂交和测序 (FISHseq) 为植入物相关感染提供了卓越的诊断. 这种独立于培养的方法可以识别额外的病原体和传统方法遗漏的生物膜形式,从而改善患者的治疗.

科学领域:

微生物学微生物学
手术病理学手术病理学
分子诊断学分子诊断学

背景情况:

植入物相关的感染带来了重大的外科手术挑战,特别是那些涉及生物膜形成微生物的感染.
传统的诊断方法,如聚合酶连锁反应和培养,难以识别致病原体并对生物膜进行分类.
准确识别病原体对于有效的治疗策略至关重要.

研究的目的:

评估光在位杂交 (FISH) 结合核酸放大和测序 (FISHseq) 对植入物相关感染的诊断价值.
将FISHseq与传统的基于培养的方法进行比较,用于识别病原体和微生物生物膜.
在手术伤口样本中绘制病原体和生物膜的空间组织图.

主要方法:

分析了来自60名怀疑患有植入物相关感染的患者的118个组织样本.
使用了经典的微生物培养以及独立于培养的FISHseq (PCR和测序的FISH).
包括从关节置换,开放缩减和内部固定以及弹射伤害的样本.

主要成果:

在60个伤口中,FISHseq在56个伤口中增加了诊断价值.
它证实了41个伤口的培养结果,并在12个伤口中确定了额外的病原体.
FISHseq将污染与真正的感染区分开来,并检测出非浮游生物的细菌形式.

结论:

除了基于培养的方法之外,FISHseq提供了重要的额外诊断信息,包括治疗相关的发现.
这种独立于培养的技术可以检测非浮游生物的细菌生命形式,有助于生物膜的表征.
FISHseq提高了复杂的植入物相关感染的诊断准确度.

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相关概念视频

FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

In-situ Hybridization

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...

Immunofluorescence Microscopy

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.