Department of Surgery-Otorhinolaryngology, Head and Neck Surgery, University of Adelaide and Flinders University, Adelaide, Australia.
Laryngoscope. 2010 Feb;120(2):427-31. doi: 10.1002/lary.20705.
OBJECTIVES/HYPOTHESIS: Biofilms are increasingly recognized as having an etiological role in chronic rhinosinusitis (CRS). Research into biofilms in CRS currently relies on microscopic imaging techniques, none of which are universally accepted. This study compares LIVE/DEAD BacLight (Invitrogen Corp., Carlsbad, CA) staining and fluorescence in situ hybridization (FISH), both utilizing the confocal scanning laser microscope (CSLM) for biofilm determination and characterization in CRS patients.
Prospective study.
Twenty CRS patients undergoing endoscopic sinus surgery were recruited for the study. Sinus mucosal tissue harvested at the time of surgery underwent both the BacLight/CSLM and FISH/CSLM protocols for biofilm determination and characterization.
Combining the results of both protocols, 18/20 (90%) patients had bacterial biofilms demonstrable on at least one modality. The high biofilm detection rate combining the two techniques suggests the prevalence of biofilms in CRS may be greater than previously reported. The protocols had equivalent results in 15/20 patients. Using the differences observed in the remaining five patients, we can highlight the most appropriate use for each technique.
BacLight/CSLM and FISH/CSLM are complementary techniques for biofilm determination and characterization. Both protocols are suited to different research areas and the selection of technique used should be based on the specific objectives of the research protocol. In this way we can utilize the advantages of each technique to facilitate effective research.
目的/假设:生物膜在慢性鼻-鼻窦炎(CRS)中的病因作用正逐渐被认识。目前对 CRS 中生物膜的研究依赖于显微镜成像技术,这些技术都没有被普遍接受。本研究比较了 LIVE/DEAD BacLight(Invitrogen Corp.,Carlsbad,CA)染色和荧光原位杂交(FISH),两者都利用共聚焦扫描激光显微镜(CSLM)来确定和描述 CRS 患者的生物膜。
前瞻性研究。
招募了 20 名接受内镜鼻窦手术的 CRS 患者进行研究。手术时采集的鼻窦黏膜组织同时进行 BacLight/CSLM 和 FISH/CSLM 方案,以确定和描述生物膜。
将两种方案的结果结合起来,18/20(90%)名患者至少有一种方法可检测到细菌生物膜。两种技术结合的高生物膜检测率表明 CRS 中的生物膜患病率可能高于先前报道。在 15/20 名患者中,两种方案的结果相同。利用其余五名患者观察到的差异,我们可以突出每种技术的最适用途。
BacLight/CSLM 和 FISH/CSLM 是生物膜确定和描述的互补技术。这两种方案都适用于不同的研究领域,选择使用的技术应基于研究方案的具体目标。这样,我们可以利用每种技术的优势,促进有效的研究。