Article(id=1220689487751987838, tenantId=1146029695717560320, journalId=1220038315760373764, issueId=1220689483859677846, articleNumber=null, orderNo=null, doi=10.12102/j.issn.1009-6493.2025.19.020, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=review-article, receivedDate=1723132800000, receivedDateStr=2024-08-09, revisedDate=1751904000000, revisedDateStr=2025-07-08, acceptedDate=null, acceptedDateStr=null, onlineDate=1768964653194, onlineDateStr=2026-01-21, pubDate=1760025600000, pubDateStr=2025-10-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768964653194, onlineIssueDateStr=2026-01-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768964653194, creator=13701087609, updateTime=1768964653194, updator=13701087609, issue=Issue{id=1220689483859677846, tenantId=1146029695717560320, journalId=1220038315760373764, year='2025', volume='39', issue='19', pageStart='3201', pageEnd='3376', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1768964652266, creator=13701087609, updateTime=1768965013333, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1220690998338961744, tenantId=1146029695717560320, journalId=1220038315760373764, issueId=1220689483859677846, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1220690998338961745, tenantId=1146029695717560320, journalId=1220038315760373764, issueId=1220689483859677846, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=3335, endPage=3341, ext={EN=ArticleExt(id=1220689488007840397, articleId=1220689487751987838, tenantId=1146029695717560320, journalId=1220038315760373764, language=EN, title=Research progress on recognition and detection of bacterial biofilms in chronic wounds, columnId=1220689485315101342, journalTitle=Chinese Nursing Research, columnName=Review Articles, runingTitle=null, highlight=null, articleAbstract=

This article reviewed the formation process of biofilms,the characteristics of wound biofilm infections,methods for identifying and detecting wound biofilms based on clinical manifestations and laboratory techniques,and promising new methods for bedside biofilm detection,with the aim of helping wound care providers to identify the presence of biofilms at an early stage accurately and to administer comprehensive biofilm management regimens.

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GUO Jinli, E⁃mail:
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对生物膜的形成过程、伤口生物膜感染的特点、基于临床表现和实验室技术识别检测伤口生物膜的方法、有望实现床旁检测生物膜的新方法进行综述,旨在为伤口护理人员及早准确识别生物膜存在并施以生物膜管理的综合方案提供帮助。

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郭锦丽,E⁃mail:
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李国庆,护士,硕士研究生在读

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李国庆,护士,硕士研究生在读

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departmentName=null, remark=2.山西医科大学第二医院)])], figs=[ArticleFig(id=1220689494181856220, tenantId=1146029695717560320, journalId=1220038315760373764, articleId=1220689487751987838, language=EN, label=Table 1, caption=

Clinical identification items for biofilm infections

, figureFileSmall=null, figureFileBig=null, tableContent=
指南与共识识别条目
生物膜感染相关指南[35]1)有感染的临床表现,如典型但常表现为低度炎症反应症状:红、肿、热、痛、功能减退,有时伴低热
2)有生物膜诱发病史(如植入性医疗器械、囊性纤维化)
3)持续感染超过7 d(非特异性,其他原因也可导致,如对所使用的抗生素产生抗药性)
4)抗生素治疗失败和感染复发
5)抗生素治疗失败的证据/病史(特别是有证据表明多个时间点的感染均由同一病原体所致)
6)抗生素治疗后全身感染症状和体征获得缓解,但停止治疗后又复发的证据
慢性不愈合伤口生物膜识别与处理的共识声明[6]1)对抗生素或防腐剂治疗抗性较高
2)抗生素或防腐剂治疗失败
3)延迟愈合
4)感染复发/恶化
5)伤口床过度潮湿与大量渗液
6)低度慢性炎症
7)轻度红斑
伤口感染与生物膜临床识别指示的国际共识[10]与国际伤口感染研究所文件[33]1)抗生素治疗失败
2)对抗生素治疗效果不佳
3)停止抗生素治疗后延迟愈合复发
4)抗菌治疗无反应
5)尽管采取了最佳的伤口管理和健康支持措施,伤口仍然延迟愈合
6)伤口渗出/湿度增加
7)低度慢性炎症
8)轻度红斑
9)肉芽生长不良/脆弱易出血的肉芽组织、肉芽组织过度生长
10)继发感染迹象
), ArticleFig(id=1220689494282519525, tenantId=1146029695717560320, journalId=1220038315760373764, articleId=1220689487751987838, language=CN, label=表1, caption=

生物膜感染临床识别条目

, figureFileSmall=null, figureFileBig=null, tableContent=
指南与共识识别条目
生物膜感染相关指南[35]1)有感染的临床表现,如典型但常表现为低度炎症反应症状:红、肿、热、痛、功能减退,有时伴低热
2)有生物膜诱发病史(如植入性医疗器械、囊性纤维化)
3)持续感染超过7 d(非特异性,其他原因也可导致,如对所使用的抗生素产生抗药性)
4)抗生素治疗失败和感染复发
5)抗生素治疗失败的证据/病史(特别是有证据表明多个时间点的感染均由同一病原体所致)
6)抗生素治疗后全身感染症状和体征获得缓解,但停止治疗后又复发的证据
慢性不愈合伤口生物膜识别与处理的共识声明[6]1)对抗生素或防腐剂治疗抗性较高
2)抗生素或防腐剂治疗失败
3)延迟愈合
4)感染复发/恶化
5)伤口床过度潮湿与大量渗液
6)低度慢性炎症
7)轻度红斑
伤口感染与生物膜临床识别指示的国际共识[10]与国际伤口感染研究所文件[33]1)抗生素治疗失败
2)对抗生素治疗效果不佳
3)停止抗生素治疗后延迟愈合复发
4)抗菌治疗无反应
5)尽管采取了最佳的伤口管理和健康支持措施,伤口仍然延迟愈合
6)伤口渗出/湿度增加
7)低度慢性炎症
8)轻度红斑
9)肉芽生长不良/脆弱易出血的肉芽组织、肉芽组织过度生长
10)继发感染迹象
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慢性伤口细菌生物膜识别与检测的研究进展
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李国庆 1 , 叶泉滟 1 , 郭锦丽 2 , 刘宏 2 , 薛媛 1 , 赵晓雨 1
护理研究 | 科研综述 2025,39(19): 3335-3341
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护理研究 | 科研综述 2025, 39(19): 3335-3341
慢性伤口细菌生物膜识别与检测的研究进展
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李国庆1, 叶泉滟1, 郭锦丽2 , 刘宏2, 薛媛1, 赵晓雨1
作者信息
  • 1.山西医科大学护理学院,山西 030001
  • 2.山西医科大学第二医院
  • 李国庆,护士,硕士研究生在读

通讯作者:

郭锦丽,E⁃mail:
Research progress on recognition and detection of bacterial biofilms in chronic wounds
Guoqing LI1, Quanyan YE1, Jinli GUO2 , Hong LIU2, Yuan XUE1, Xiaoyu ZHAO1
Affiliations
  • 1.Nursing College of Shanxi Medical University, Shanxi 030001 China
  • 2.Second Hospital of Shanxi Medical University
出版时间: 2025-10-10 doi: 10.12102/j.issn.1009-6493.2025.19.020
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对生物膜的形成过程、伤口生物膜感染的特点、基于临床表现和实验室技术识别检测伤口生物膜的方法、有望实现床旁检测生物膜的新方法进行综述,旨在为伤口护理人员及早准确识别生物膜存在并施以生物膜管理的综合方案提供帮助。

慢性伤口  /  细菌生物膜  /  生物膜  /  床旁检测  /  综述

This article reviewed the formation process of biofilms,the characteristics of wound biofilm infections,methods for identifying and detecting wound biofilms based on clinical manifestations and laboratory techniques,and promising new methods for bedside biofilm detection,with the aim of helping wound care providers to identify the presence of biofilms at an early stage accurately and to administer comprehensive biofilm management regimens.

chronic wounds  /  bacterial biofilms  /  biofilm  /  point⁃of⁃care testing  /  review
李国庆, 叶泉滟, 郭锦丽, 刘宏, 薛媛, 赵晓雨. 慢性伤口细菌生物膜识别与检测的研究进展. 护理研究, 2025 , 39 (19) : 3335 -3341 . DOI: 10.12102/j.issn.1009-6493.2025.19.020
Guoqing LI, Quanyan YE, Jinli GUO, Hong LIU, Yuan XUE, Xiaoyu ZHAO. Research progress on recognition and detection of bacterial biofilms in chronic wounds[J]. Chinese Nursing Research, 2025 , 39 (19) : 3335 -3341 . DOI: 10.12102/j.issn.1009-6493.2025.19.020
慢性伤口已成为全球性的卫生健康问题,发达国家慢性伤口病人占其总人口的1%~2%[1],我国每年约3 000万例慢性伤口病人需要护理[2],人均治疗费用高,给病人和卫生保健系统造成巨大负担。细菌生物膜被证实是慢性伤口不愈合的重要原因,其会诱导伤口局部持续炎症反应、抑制伤口成纤维细胞发育并降低伤口部位抗拉强度等,使慢性伤口迁延不愈甚至愈合后又复发[3]。生物膜在慢性伤口中流行率极高,研究显示,近80%的慢性伤口存在生物膜[4],在特定的伤口类型,如糖尿病足溃疡中,其流行率甚至可达100%[5]。鉴于生物膜对慢性伤口的影响,相关共识指南提出了标准处理措施——基于生物膜的伤口护理(biofilm⁃based wound care,BBWC)[6]。其中,准确识别生物膜感染是第一步,也是实施基于生物膜伤口护理的前提。然而,由于尚缺乏生物膜诊断的“金标准”[7],并且目前的技术主要用于诊断伤口浮游细菌感染,难以准确识别细菌生物膜,导致医护人员判定生物膜感染并施行基于生物膜的伤口护理存在较大困难。基于此,本研究就慢性伤口细菌生物膜概述、细菌生物膜临床识别、实验室检测进行综述,并介绍有望实现床旁检测生物膜的新技术,旨在为伤口医护人员准确识别并清除细菌生物膜提供帮助。
在任一感染部位,微生物主要以分散态的浮游细菌和聚集态的细菌生物膜2种不同的表型存在[8]。细菌生物膜被定义为附着在活性或惰性物质表面的具有遗传多样性和可变基因表型的结构化复杂微生物群落,包裹于其产生的由多糖、蛋白质、脂质和胞外DNA等组成的胞外聚合物(extracellular polymeric substances,EPS)中[910],导致慢性感染产生并持续存在。
细菌生物膜形成具有复杂性、动态性、循环性,涉及细菌生理特性变化并遵循既定的生长周期,包含如下阶段[1114]。1)可逆的粘附阶段:伤口床富有营养物质,浮游细菌通过弱相互作用附着于其上,此时作用力微弱,粘附是可逆的。随着粘附发生,生物膜细菌的表型逐渐有别于浮游形态。2)不可逆的定植阶段:细菌进一步通过自身结构或分泌某些物质等不可逆附着于伤口表面,成功附着后,开始启动基因表达,逐步产生保护性胞外聚合物,以强化附着并为自身提供结构和生化支持。同种细菌相互吸引形成微菌落,初步形成生物膜结构。3)发育成熟阶段:该阶段涉及不同种类细菌的协同能力,细菌增殖聚集形成稳定成熟的三维立体菌落结构,具体形态取决于细菌种类。成熟生物膜的最大平均厚度50~100 mm,体积构成以胞外聚合物为主(占90%),细菌仅占10%。特殊的结构造就生物膜独特的特性,至此,生物膜细菌的基因表达与浮游细菌截然不同。4)脱离与播散阶段:成熟生物膜中细菌数量不断增加、代谢废物不断累积,而营养物质却逐渐减少,细菌在多种因素作用下单个或成团从生物膜中脱离,播散定植至伤口局部或较远环境,脱离的细菌重新恢复浮游细菌表型,重新开始生物膜的下一循环周期。在生物膜形成的动态过程中,可逆的黏附阶段为始,但脱离与播散阶段却并不是终点,而是下一生物膜循环形成的开始,也是造成临床伤口感染反复发生的主要原因之一。
慢性伤口生物膜菌群种类繁多[15],通常由占主导地位的几种细菌及上百种其他不同类微生物构成,多微生物产生可协同造成宿主损伤[16]。综合现有文献,慢性伤口生物膜中常见的优势菌群为金黄色葡萄球菌、铜绿假单胞菌、粪肠球菌、肺炎克雷伯菌、鲍曼不动杆菌和肠杆菌属等,最常见的为金黄色葡萄球菌[17]。分子技术促进了对慢性伤口微生物群和生物膜微生物群的认识。Wolcott等[18]使用16s rDNA测序技术分析了2 963例病人的各类慢性伤口(压力性损伤、糖尿病足溃疡、静脉溃疡等),发现葡萄球菌属和假单胞菌属是最常见的菌属,同时强调了厌氧菌的高流行率,并且观察到伤口类型与细菌组成之间似乎并无明显关联。迄今为止,研究者的关注点主要集中于生物膜中的各种细菌病原体,但与此同时,真菌(尤其是念珠菌)在慢性伤口生物膜中的重要作用也逐渐被发现[1920]
研究表明,生物膜细菌的耐药性是浮游细菌的10~1 000倍[21]。因此,尽管给予恰当抗菌治疗,生物膜感染仍可能长期存在,并可能在治疗结束后导致感染复发[9,22]。其抗菌耐受性增强的机制可归纳如下[2328]
胞外聚合物作为生物膜细菌的机械屏障,可减缓或抑制药物和免疫细胞向生物膜中扩散,保护内部细菌免受药物和宿主免疫系统的杀伤与清除,且免疫系统在对抗成熟生物膜相关感染时往往效果不佳。
生物膜内存在氧气梯度,表面的氧浓度最高,向内逐渐下降,在中心几乎形成厌氧环境。低氧条件有利于生物膜形成,而常氧条件则产生抑制作用。生物膜内的细菌根据自身对缺氧的耐受程度和新陈代谢速率有序排列,位于表面的细菌代谢活跃,位于深层的细菌细胞生长十分缓慢或处于休眠状态,进入存活但不可培养(viable but non⁃culturable,VBNC)状态,往往需要更长时间抗生素治疗才能杀灭。
群体感应是指菌体自身产生化学信号并且感知其相应信号浓度变化,进行细菌种间或种内信息交流,从而调节细菌群体行为的一种特殊调控系统,抗生素耐药性也被证实与群体感应密切相关,因群体感应可诱导生物膜细菌进入存活但不可培养状态。
要彻底根除伤口生物膜,关键性举措是破坏生物膜胞外聚合物[2930]。清创是破坏生物膜胞外聚合物的最关键手段[31]。然而,生物膜在伤口中并非均匀分布且多数生物膜位于伤口床深层组织(平均深度50~70 μm)[6],加之临床尚未发现能可视化伤口生物膜分布图的手段,既无法明确伤口清创范围、程度,也无从判断清创前后生物膜去除效果,导致生物膜残留不可避免。研究表明,残留的生物膜细菌可在24 h内迅速重建生物膜结构,并且重建的生物膜会变得更难以去除[32]
准确识别伤口生物膜对于制定伤口护理方案至关重要,但伤口生物膜临床识别仍具有挑战性。活性组织生物膜体积仅4~200 μm,缺乏宏观上肉眼明辨的特征[6],并且伤口生物膜还可嵌入腐肉、碎屑、坏死组织,甚至伤口敷料中,既可被覆于伤口表面,又可深埋于伤口深部组织,进一步增加了识别难度[33]。早期观点认为,苍白的伤口床、黏稠的伤口床表面物质等可视为伤口生物膜存在[34],但有证据表明,肉眼完全无法识别伤口生物膜结构[33]。因此,经大量实验研究及临床实践,相关文件总结发布了间接识别伤口生物膜的“临床线索”。2014年,欧洲临床微生物学和传染病学会发布了第1份生物膜感染相关指南[35],初步明确了生物膜感染的一般特征,适用于绝大多数生物膜相关感染的临床判定,但对于识别伤口生物膜感染特异性欠佳、指导价值有限。之后,2017年[6]与2019年[10]的共识声明及2022国际感染研究所文件等[33]逐步增加了伤口局部临床表现条目,如伤口大量渗液、肉芽组织生长不良、脆弱易出血的肉芽组织、肉芽组织过度生长等,更具象化、更利于伤口护理人员识别生物膜感染。详见表1。伤口生物膜感染的临床指征可在一定程度上帮助伤口护理人员高度怀疑或初步判定伤口生物膜存在,从而决定是否采取针对性处理措施。然而,其中某些指标的准确识别高度依赖判定者自身的临床经验,存在主观性,且实验室研究证明,很多肉眼看似正常的伤口实际却存在生物膜[36]。因此,仍需依赖更精确、客观的手段进一步检测伤口生物膜,例如实验室技术。
伤口生物膜的实验室检测主要侧重于2个方面:1)生物膜细菌鉴定;2)检测生物膜胞外聚合物基质。
基于细菌培养的技术具有成熟完善的操作规程,是目前临床常规使用的检测感染致病菌的标准方法。彻底清洁清创后的伤口深部组织标本是揭示伤口生物膜最可靠的样本[35],鉴于伤口生物膜的分布特点,建议从伤口浅层和深层获取多个组织样本以提高检测敏感性。在无法获取组织样本时,Levine伤口拭子技术可作为替代方法,但需注意避免正常菌群污染导致的假阳性结果[37]。尽管标准培养技术在临床实践中应用广泛,但其检测生物膜细菌仍具有挑战性。研究显示,传统培养方法仅能识别慢性伤口中1%的细菌[34]。首先,细菌状态影响检出率[38],标准培养主要针对存活可培养的浮游细菌,但浮游细菌的种类可能与形成生物膜的细菌不同。其次,慢性伤口生物膜包含多种微生物,多细菌共存使得细菌的可培养性受到其他细菌的影响,增加培养难度[16];此外,生物膜中大量细菌处于存活但不可培养状态,正常培养条件下难以形成菌落,导致检测结果不准确[26]。最后,标准培养方法需要花费数天时间才能报告检验结果,在此期间,生物膜可能已扩散到其他部位[39]。因此,基于传统细菌培养方法评估生物膜细菌可能不够充分、不够准确且耗时较长。
虽然目前尚无伤口生物膜诊断的“金标准”,但欧洲临床微生物学和传染病学会生物膜研究小组指出,扫描电子显微镜(scanning electron microscope,SEM)和激光扫描共聚焦显微镜(confocal laser scanning microscope,CLSM)是细菌生物膜最可靠和客观的诊断手段[6]。胞外聚合物由多糖、蛋白质、脂类等组成,确定并量化胞外聚合物成分有助于研究人员明晰生物膜的成熟度和复杂性。
扫描电子显微镜是一种基于电子表面散射和吸收的技术,适用于观察生物膜的表面结构,结合成像软件可评估生物膜的三维数据[40]。对伤口病原体和生物膜鉴定具有高度敏感性和特异性,其可以发现未出现明显感染迹象伤口的细菌生物膜[35],通过扫描电子显微镜,可观察到没有鞭毛的铜绿假单胞菌包裹在无炎症迹象的伤口细胞外基质中[41]。扫描电子显微镜还可用于评估抗生物膜化合物的功效及为定量研究结果提供定性支持等[42]。其不足之处是缺乏垂直分辨率、样品制备过程烦琐耗时且可能会破坏样品结构或造成伪影而影响观察。
激光扫描共聚焦显微镜是目前生物膜可视化和量化的首选方法[16],有助于了解生物膜内部结构,可于生物膜不同深度平面采集多重图像,形成不同景深的高分辨率图像,配合图像分析软件,可呈现生物膜空间结构并量化生物膜的结构参数,如生物膜的生物体积、厚度和粗糙度等[4344]。此外,结合针对生物膜成分可视化的分子探针和荧光染料,激光扫描共聚焦显微镜可识别生物膜多糖、蛋白质和脂质等大分子,从而明确生物膜的不同物质组成、确定参与生物膜形成的关键因素及观察细胞外基质和生物膜的空间关系等[4546],为临床制定干预措施提供参考。
尽管电子显微镜可提供准确、可靠的结果,但该检测方法需要昂贵、特殊的设备支持和专业人员操作,目前主要用于科研,在临床环境中难以常规开展。
床旁检测指在现场实时取样、实时分析,免去繁杂的实验室处理程序,快速获得检验结果的方法,具有设备小型便携化、操作简便化、报告即时化和结果客观化等特点,已被用于进行血糖、妊娠监测以及感染性疾病筛查等领域[4748]。鉴于伤口生物膜临床识别的主观性、实验室检测的耗时性、复杂性、不可及性等,床旁(即时)检测技术在伤口生物膜感染检测中天然具有广阔应用前景。目前已有多项相关临床研究,主要包括检测生物膜细菌和检测生物膜胞外聚合物2种方法。
识别、减少并消除细菌负荷是降低伤口感染风险、促进伤口愈合的基础[37]。传统观念认为,细菌量>105 cfu/g时会发生感染并使伤口愈合延迟[49]。但Armstrong等[50]提出了新的术语——慢性抑制性细菌负荷(chronic inhibitory bacterial load,CIBL),即>104 cfu/g的高细菌负荷状态,这一临床相关水平的细菌负荷并不会使伤口表现出明显的感染迹象,但却会损伤组织并抑制伤口愈合进程。目前多依据微生物定量培养及临床症状体征检查表评估伤口细菌负荷,但微生物定量培养程序复杂、结果报告滞后,临床症状体征检查表的灵敏度又极低,无法发现超过80%的高细菌负荷伤口[51]。因此,亟须准确、可靠并能立即报告结果的评估伤口高细菌负荷的工具。
MolecuLight i∶X细菌荧光成像设备填补了这一领域的空白,其原理是在距伤口床8~12 cm处发出405 nm紫光照射于伤口,激发伤口组织发出不同颜色荧光,即刻呈现伤口CIBL区域的位置信息[52]。成像的荧光信号显示在该设备的数字触摸屏上,查看荧光图像,皮肤成分(如胶原蛋白、角蛋白)产生绿色荧光信号(阴性),慢性抑制性细菌负荷区域产生红色和/或青色荧光信号(阳性)[53]。具体来说,红色荧光(也可为粉红色、胭脂红或黄色)代表超过28种(革兰阳性菌、革兰阴性菌、需氧菌、厌氧菌)可产生卟啉的伤口感染常见细菌(如葡萄球菌属、变形杆菌属、克雷伯氏菌属、拟杆菌属等),青色荧光则代表产生吡咯烷酮的铜绿假单胞菌[54]。荧光成像可准确、实时地发现伤口床、伤口周围皮肤甚至伤口周围皮肤以外的慢性抑制性细菌负荷区域,阳性预测值高达95%~100%[51,5556]。Lopez等[57]研究表明,该设备不仅可检测到伤口浮游细菌的荧光信号,还可穿透生物膜胞外聚合物检测生物膜内部细菌的荧光信号,有助于指导生物膜细菌的可视化清创,加速伤口愈合。Rahma等[58]报道了首个评估在糖尿病足溃疡标准护理中加入细菌荧光成像的随机对照试验,结果显示,荧光成像组的12周伤口愈合率(45%)比对照组(22%)提高了1倍,伤口愈合率的显著提升归功于荧光成像指导下的针对性的彻底清创措施及清创效果的即时反馈。Price[59]回顾性研究显示,常规使用细菌荧光成像使糖尿病足病人的截肢率降低了11%。迄今为止,荧光成像已积累了大量临床数据,既验证了其慢性抑制性细菌负荷区域的诊断准确性,又证明了其在改善伤口护理计划方面的实用性,为清创[58,60]、微生物分析取样位置选择[6162]以及是否需要升级处理措施或进行进一步检查等方面决策提供积极支持[60]
细菌荧光成像具有及时、准确、无创等优势,显著提升了病人的伤口治疗效果,改善了伤口护理实践。但其也有一些局限性:1)其机制主要是基于检测生物膜内的细菌荧光信号,而不是生物膜本身,因此当生物膜内的细菌量不足以达到其检测阈值时,可能会导致假阴性结果;2)其无法检测到不产生卟啉的细菌,如链球菌属、肠球菌属等,尽管这些细菌在慢性伤口中并不常见;3)其可以检测但无法区分生物膜细菌与浮游细菌;4)其无法明确细菌种类及抗生素药物敏感或耐药情况等。
目前尚无可明确伤口生物膜确切位置分布的技术,因此及时采取措施精准清除伤口生物膜具有挑战性。伤口印迹法是一种新兴的床旁检测技术,首先在伤口表面贴敷可吸收伤口渗液并固定蛋白质、多糖等生物分子的带阳离子电荷的膜,待生物膜胞外聚合物成分吸附到膜上后,使用阳离子染料染色进行下一步分析[63]。胞外聚合物占生物膜的90%,细菌仅占10%,因此不同于细菌荧光成像,伤口印迹法直接检测生物膜胞外聚合物含量最高的多糖成分,结果更稳定,灵敏度更高。该方法不仅可以准确检测伤口生物膜,还可以提供伤口生物膜相关定性信息,如生物膜的二维位置分布和伤口愈合的预后。Azeredo等[42]使用伤口印迹法前瞻性检测压力性损伤伤口生物膜形成,使用硝酸纤维素膜吸收多糖,用钌红进行染色,结果显示,超过80%的阳性染色伤口在1周后愈合状况无明显改善或伤口腐肉至少增加10%。因此,伤口印迹法不但可以用于在临床环境中检测细菌生物膜,其阳性结果还可作为压力性损伤延迟愈合的指征,指导伤口护理人员提前做出干预。但Azeredo等的方法只能提阳性或阴性结果,而且样本染色处理等时间较长(>3.0 min),难以满足即时检测的要求。Wu等[64]对此方法进行了改良,全部操作过程可在2 min左右完成:首先将尼龙膜贴覆于伤口床10 s,之后在阳离子洗涤剂中浸泡30 s,用阿尔新兰进行染色,再在前述阳离子洗涤剂中浸泡并振荡30 s,风干后,生物膜多糖成分在膜上呈蓝色。改良方法除可明确生物膜在伤口的分布外,还可可视化分级生物膜感染严重程度并预测90 d的伤口愈合情况。染色分级共分4级,0级为阴性,膜上无可见的蓝色,1~3级为阳性,膜上蓝色强度依次递增。90 d的随访结果显示,初始染色为2级或3级的病人中,0%的伤口在30 d内愈合,42.9%~50.0%的伤口在30~90 d愈合,50%~57.1%的伤口在90 d随访时仍未愈合。而在初始染色为0级或1级的病人中,90 d内伤口愈合率为100%,77.8%的0级和14.3%的1级伤口在30 d内愈合,22.2%的0级和85.7%的1级伤口在30~90 d愈合。
伤口印迹技术是一种非侵入性、即时、精确、客观的生物膜检测方法,适用于大多数临床环境,将来有望实现生物膜的床旁检测,从而促进基于生物膜的伤口护理实施,优化伤口管理。其提供的生物膜分布信息可指导进行不损伤周围正常组织的针对性清创,染色分级可直接理解为生物膜感染的严重程度,可预测伤口愈合进展。但伤口印迹技术也具有一定的局限性:1)无法明确细菌种类及抗生素药物敏感或耐药情况等;2)在干性坏疽或固着焦痂覆盖的伤口中灵敏度不佳(难以收集足够样本量的伤口渗液)。
人们越来越意识到识别、检测并根除伤口生物膜对促进慢性伤口愈合的积极作用。一项文献计量学研究表明,检测伤口生物膜感染一直是研究人员关注的热点领域[65]。目前,伤口护理人员首先依靠临床线索初步怀疑或判定伤口生物膜感染,但该方法主观性较强、准确性欠佳。实验室技术可精确诊断生物膜感染,但其代价高昂、耗时长且无法在临床环境中常规使用。新技术的出现有望实现生物膜的床旁精确检测,指导实施以生物膜为导向的临床治疗。MolecuLight i∶X细菌荧光成像设备可快速、准确识别包括金黄色葡萄球菌和铜绿假单胞菌在内的高生物膜形成倾向性细菌的慢性抑制性细菌负荷区域荧光信号,灵敏度高,可在诊疗过程中重复进行,提供处理措施的反馈信息,帮助伤口护理人员减少伤口细菌负荷,促进伤口愈合。伤口印迹法可以非侵入性的方式直观绘制生物膜在伤口表面的分布图,量化生物膜感染严重程度,操作简便,结果报告及时,可指导伤口护理人员采取针对性的伤口清创,提高后续抗菌处理措施效果,改善伤口愈合状况。但这2种技术目前主要用于指导采取针对性清创措施,无法获得有关药物敏感性的信息,这对于处理播散性甚至全身性严重感染可能存在不足,未来可考虑与基于培养的技术结合使用。此外,还需要更大范围、更广泛区域、更高质量的随机对照研究进一步验证这2种技术的临床适用性、经济性及普及性等。
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2025年第39卷第19期
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doi: 10.12102/j.issn.1009-6493.2025.19.020
  • 接收时间:2024-08-09
  • 首发时间:2026-01-21
  • 出版时间:2025-10-10
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  • 收稿日期:2024-08-09
  • 修回日期:2025-07-08
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    1.山西医科大学护理学院,山西 030001
    2.山西医科大学第二医院

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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