16S rRNA 测序鉴定1株条件致病性人苍白杆菌

目的：探索一种基于16S rRNA 基因的细菌快速鉴定方法,为临床未知病原菌的诊断及治疗提供科学依据。方法对临床患者的痰标本分离培养纯菌落,直接以菌液为模板,以通用引物 PCR 扩增未知菌的16S rRNA 基因片段,产物直接测序。将测序结果进行 BLAST 比对,根据序列同源性鉴定病原细菌。结果未知病原菌经本实验鉴定为人苍白杆菌,经 ABI 细菌快速鉴定板条检测,确认结果一致。结论该研究简化了临床标本未知病原菌分离培养鉴定的步骤,建立了一种利用16S rRNA 基因扩增快速鉴定病原菌的简便方法。     

23S rRNA基因在常见病原菌鉴定中的应用

目的建立基于23S rRNA基因的一种常见病原菌菌种鉴定的分子生物学方法。方法使用地高辛标记的通用引物扩增常见病原菌的23S rRNA基因，并在尼龙膜上固定根据扩增产物序列设计的菌种特异性探针，根据反向杂交结果判断病原菌种类。选取临床分离获得的菌株验证该方法的有效性。结果通用引物可特异性扩增细菌23S rRNA。应用该方法可鉴定金黄色葡萄球菌、凝固酶阴性葡萄球菌、粪肠球菌、屎肠球菌、肺炎链球菌、大肠埃希菌、肺炎克雷伯菌、奇异变形杆菌、阴沟肠杆菌、鲍曼不动杆菌、绿脓假单胞菌、嗜麦芽窄食单胞菌和洋葱伯克霍尔德菌。检测临床标本结果显示，与常规培养方法的符合率为99％。结论该方法可用于常见病原菌的鉴定。     

16S rRNA通用引物在血小板制品细菌污染检测中的应用

目的评价16S rRNA通用引物在快速检出血小板制品中污染细菌的实用性,寻找新的能够快速、准确地检出血小板中污染细菌的方法。方法用分子生物学方法提取样本中细菌DNA,用设计合成的16S rRNA和16S—23S rRNA基因区间引物进行扩增,产物经HaeⅢ酶切,酶切片段与血小板污染常见菌的酶切图谱对比,确定有无污染及污染菌种类。结果16S rRNA通用引物法可以在4h内完成全部检测,其中在保存24h的血小板标本中未检出污染菌,保存48h的血小板标本中有2份分别检出了金黄色葡萄球菌和枯草芽孢杆菌,与全自动细菌培养仪的结果一致。结论16S rRNA通用引物是一种快速、准确检出血小板制品中污染细菌的方法,结果可靠,具有较高的临床应用价值。     

细菌性肺炎快速诊断基因芯片的初步构建

目的：根据细菌16 S rRNA基因特点设计常见病原菌的特异性探针,采用酶显色技术构建基因芯片,探讨其临床应用的可能性。方法选取肺炎链球菌、流感嗜血杆菌及铜绿假单胞菌等8种细菌性肺炎常见的病原菌的标准菌株作为研究对象,并选择12份患者的痰液标本进行检测。在16 S rRNA基因保守区设计 PCR反应的通用引物及革兰阳性菌、革兰阴性菌的通用探针,利用可变区的差异设计合成特异性探针,构建基因芯片。利用细菌16S rRNA基因设计的PCR通用引物进行扩增,所有8种细菌均获得350 bp的扩增产物。以地高辛标记特异性探针,构建完成可用于8种常见病原菌检测的基因芯片,结果8种标准菌株基因芯片检测均取得了预期效果,对12份痰标本中常规培养阳性7份,其对应芯片检测结果均成阳性,5份常规培养阴性的标本中,芯片结果提示阳性的有3份,其中1份为嗜肺军团菌,2份为使用抗生素后的患者标本。结论设计合成的 PCR通用引物对扩增细菌的16 S rRNA基因具有较高的特异性及灵敏度。构建的基因芯片可用于常见细菌性肺炎病原菌的检测鉴定,且对抗生素使用后的临床标本及苛养菌有一定的诊断价值。本研究所获得基因芯片对于细菌性肺炎的检测具有简单、快速、特异性及敏感性高的特点。     

用16S rRNA基因克隆文库研究腹泻粪便中菌群分布

目的 建立16S rRNA基因克隆文库分析菌群的方法,用于临床标本菌群分布的检测.方法 临床标本直接提取核酸,用16S rRNA基因的通用引物进行PCR扩增、纯化、连接、克隆和测序,建立16s rRNA基因克隆文库,序列与数据库进行比对分析.结果 通过16S rRNA基因克隆文库分析,腹泻标本中检出4种细菌,脆弱拟杆菌为绝对优势菌,占91%;腹泻恢复后的粪便标本中菌群呈多样性,检出12种确定种属的细菌,其中脆弱拟杆菌占14%.结论 16S rRNA基因克隆文库是一种较好地研究粪便标本菌群的方法,可直接进行粪便标本的菌群分析和研究各种细菌在腹泻中的作用.     

急性淋巴细胞白血病患者血液中草螺菌的鉴定

目的 分离鉴定源于急性淋巴细胞白血病（ALL）患者血液中的未知细菌.方法 采用常规及VITEK32微生物GNI鉴定卡鉴定细菌,K-B法进行药敏试验,PCR法扩增细菌的16S rRNA基因,通过测序并与GenBank中相关序列进行比对.结果 常规生化反应和微生物自动化鉴定系统不能鉴定.16S rRNA PCR扩增产物经测序与GenBank中序列比对,发现与草螺菌属有99%同源性.结论 来源于ALL患者血液中的未知细菌是草螺菌.     

荧光定量PCR/熔解曲线分析法快速鉴定葡萄球菌菌种

目的：建立一种基于葡萄球菌16～23s rRNA间隔区序列特征的荧光定量PCR／熔解曲线分析鉴定方法，以快速鉴定葡萄球菌菌种。方法：在葡萄球菌16s rRNA 3′端和23s rRNA5′端分别设计上、下游引物．用荧光定量PCR方法扩增各个实验菌株的16～23s rRNA间隔区序列。然后分析各扩增产物的熔解曲线．以确定各个菌种熔解曲线特征，并以此为依据对15株葡萄球菌临床分离株进行鉴定。结果：设计的引物对成功地扩增了葡萄球菌所有实验菌株，对扩增产物的熔解曲线分析可初步鉴定金黄色葡萄球菌、表皮葡萄球菌和溶血葡萄球菌。以VITEK2鉴定结果为标准，荧光定量PCR／熔解曲线分析法鉴定菌种的准确率为67％（10／15）．整个实验可在1h内完成。结论：荧光定量PCR／熔解曲线分析法有望成为临床实验室快速鉴定葡萄球菌菌种的新方法。[著者文摘]     

临床常见病原菌16S rDNA单链构象多态性分析

目的 原核生物的rRNA基因位点具有高度的保守性,同时不同种属细菌之间又存在相对稳定的变异性,因此可被用于细菌的分类和鉴定.本研究试图利用16S rRNA基因结合分子生物学方法,达到快速鉴定临床常见病原菌的目的.方法 收集临床常见各种病原菌333株,筛选2对通用引物在5′端用不同荧光标记,然后对细菌基因组DNA进行聚合酶链反应（PCR）扩增,产物在测序仪上通过毛细管电泳-单链构象多态性（CE-SSCP）分析,通过不同细菌的峰谱不同达到快速鉴定病原菌目的.结果 所有受试细菌经过CE-SSCP分析后都有各自不同的峰谱出现,相互之间容易区别.结论 利用PCR-CE-SSCP作为病原菌鉴定手段,高效、准确,较传统方法省时,而且灵敏度较高,是一种很有应用前景的细菌快速鉴定方法.     

假性动脉瘤患者感染厌氧消化链球菌的鉴定

目的 分离鉴定源于假性动脉瘤破裂继发急性坏疽患者感染组织中的未知细菌.方法 PCR法扩增细菌的16SrRNA基因,通过测序并与GenBank中相关序列进行比对.结果 16S rRNA PCR扩增产物经测序与GenBank中序列比对,发现与厌氧消化链球菌有99％同源性.结论来源于该患者组织中的未知细菌是厌氧消化链球菌.     

应用基因芯片快速检测临床常见细菌

目的 应用基因芯片对临床常见的8种致病细菌进行检测.方法 选取8种临床常见的致病细菌,包括金黄色葡萄球菌、铜绿假单胞菌、肺炎克雷伯菌、大肠杆菌、奇异变形杆菌、产气肠杆菌、荧光假单胞菌、宋内志贺菌.以16S rRNA基因为目的基因,白行设计通用引物系列扩增目的片段,针对高变区域设计探针,建立基因芯片检测体系,并对所选细菌...     

Live a legacy or live a lie: a choice

This article is about the result of a choice to Live a Legacy or Live a Lie. The choice led to the development and implementation of a clinical practice model (CPM) designed to create a healthy, healing integrated practice culture. The focus is on the foundation of the model that is a unique ongoing process called the Core Belief Review. The purpose of the review is to uncover those things that matter most for both those who give and receive care. The ongoing open communication process provides insights and truths about reality and a sense of direction related to the nature of the work necessary to create and sustain the best places to give and receive care. The results of the review feedback from 2500 providers and recipients of care are correlated to the actions taken to address the complexities of the point-of-care reality and the clinical outcomes reached as a result of collaboration and lessons learned within an International Consortium.     

Intensive care use in a developing country: a comparison between a Tunisian and a French unit

Objectives: To compare the variations in intensive care (ICU) outcome in relation to variations in resources utilization and costs between a developed and a developing country with different medical and economical conditions. Design and setting: Prospective comparison between a 26-bed French ICU and an 8-bed Tunisian ICU, both in university hospitals. Patients: Four hundred thirty and 534 consecutive admissions, respectively, in the French and Tunisian ICUs. Measurements: We prospectively recorded demographic, physiologic, and treatment information for all patients, and collected data on the two ICU structures and facilities. Costs and ICU outcome were compared in the overall population, in three groups of severity indexes and among selected diagnostic groups. Results: Tunisian patients were significantly younger, were in better health previously and were less severely ill at ICU admission (p < 0.01). French patients had a lower overall mortality rate (17.2 vs 22.5 %; p < 0.01) and received more treatment (p < 0.01). In the low severity range, the outcome and costs were similar in the two countries. In the highest severity range, Tunisian and French patients had similar mortality rates, while the former received less therapy throughout their ICU stays (p < 0.05). Conversely, in the mid-range of severity, mortality was higher among Tunisian patients, and a difference in management was identified in COPD patients. Conclusion: Although the Tunisian ICU might appear more cost-effective than the French one in the highest severity group of patients, most of this difference appeared in relation to shorter lengths of ICU stay, and a poorer efficiency and cost-effectiveness was suggested in the mid-range severity group. Differences in economical constraints may partly explain differences in ICU performances. These results indicate where resource allocation could be directed to improve the efficiency of ICU care. Received: 15 December 1997 Accepted: 8 July 1998     

Caring for a patient with a urostomy in a community setting

Around 7500 people in the UK have a urostomy. A urostomy is normally performed if a person has bladder cancer, congenital bladder abnormalities. Many people who have a urostomy have long term conditions and may require the help and support of community nurses. This article examines common complications of urostomy including stomal complications, urinary tract infections and dermal complications. Although stoma complications are common and can affect quality of life, many people with a stoma tend not to seek help. Community nurses can provide care and support to optimize stoma management and enable the person with a urostomy to enjoy the best possible quality of life.