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1.
MALDI-TOF-MS在病原微生物鉴定中的研究进展   总被引:5,自引:0,他引:5  
基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)是鉴定多种致病性细菌的快速、可靠的方法,具有较好的稳定性和可重复性,在快速和准确性方面的总体表现明显好于传统的细菌生化鉴定方法。适合于一些致病菌的快速、高通量的检测和鉴定。综述MALDI-TOF-MS技术在普通病原菌、多血清型病原菌、非发酵性细菌,以及植物病原菌等病原微生物鉴定方面的最新研究进展。  相似文献   

2.
目前建立了多种对微生态细菌鉴定分型的DNA分析技术,这些方法使细菌分类的水平不断提高,由最初的区分不同属、种的细菌发展到同种不同株细菌分类区分。本文对这些遗传学方法的原理、特性进行了简单的介绍和比较。  相似文献   

3.
目的研究基质辅助激光解析电离飞行时间质谱(Matrix-Assisted Laser Desorption Ionization-Time of Flight MassSpectrometry,MALDI-TOF-MS)用于快速检测鉴定临床分离的酵母菌的可行性。方法应用Bruker MALDI-TOF-MS和VITEK 2-compact系统分别鉴定150株临床分离的酵母菌,结果不一致的菌株通过基因序列测定来鉴定。结果 MALDI-TOF-MS快速准确鉴定出了150株临床酵母菌,鉴定符合率在属水平上为100%,种水平上为94%。结论基于MALDI-TOF-MS鉴定方法具有很好的可重复性和准确性,并且其检测成本较低,实验准备时间很短,MALDI-TOF-MS可以用于临床分离的酵母菌的快速鉴定。  相似文献   

4.
【目的】比较16S rRNA和recA、groEL基因部分序列用于乳酸乳球菌乳酸亚种和乳脂亚种分类鉴定的效果。【方法】对已鉴定的8株分离自传统发酵乳的乳酸乳球菌, 选取recA和groEL基因片段, 通过PCR扩增、测序, 将测序得到的序列比对后构建系统发育树, 并与16S rRNA基因序列分析技术进行比较。【结果】比较分析不同菌株16S rRNA和recA、groEL基因的亲缘关系, recA、groEL基因可以准确地完成乳酸乳球菌乳酸亚种和乳脂亚种的区分和鉴定。【结论】recA和groEL基因序列分析可以实现乳酸乳球菌乳酸亚种和乳脂亚种的区分, 因其具有快速、准确、稳定的特点, 可适合于乳酸乳球菌乳酸亚种和乳脂亚种间的快速分类鉴定。  相似文献   

5.
运用基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of-flight mass spectrometry,MALDI-TOF-MS)技术快速鉴定食品中空肠弯曲菌。通过对该方法的样品前处理的选择、稳定性、特异性等方面进行研究,确定了方法的可行性。结果表明,MALDI-TOFMS鉴定方法准确,快速,稳定性好,能够准确快速地区分弯曲菌近缘菌株,并与VITEK生化鉴定结果高度一致,在准确性和时效性上均有较大提高。MALDI-TOF-MS方法适用于食品中空肠弯曲菌的快速筛检。  相似文献   

6.
目的探讨Vitek-AMS对临床细菌鉴定的应用价值。方法对玉溪市人民医院1999年至2008年临床分离11 537株细菌(临床株)和省、部级临床检验中心下发微生物学室间质量评价鉴定菌种(参考株)的Vitek-AMS鉴定结果作对比分析。结果 11 537株临床分离菌中,不能鉴定细菌8株(0.07%);除外传染病因子,鉴定到属细菌114株(1.62%),鉴定到种(含亚种、物生型)细菌6 908株(98.38%)。共有64属193种。细菌类型的分布革兰阴性杆菌革兰阳性球菌酵母菌革兰阳性杆菌厌氧菌革兰阴性球菌。用参考菌种作比较,Vitek鉴定种的符合率为83.08%(54/65),属的符合率为98.46%(64/65);其中革兰阴性杆菌符合率最高(100%,24/24),酵母菌类符合率最低(66.67%,14/21)。7种鉴定卡菌种的阳性检出率平均为56.80%(234/412),其中GPI卡最高(100%),NHI卡最低(13.33%);但应用机会最多是GNI+卡。Vitek-AMS检测葡萄球菌产β-内酰胺酶阳性率为88.89%,大肠埃希菌和肺炎克雷伯菌产ESBLs阳性率分别为59.74%和32.20%。结论 Vitek-AMS的应用为临床细菌学检验提供了一种高效、快速、可靠的实验方法;但对于个别菌种、细菌酶的检测必要时应以参考方法确认。  相似文献   

7.
温郁金内生拮抗细菌B-11的分离及其抑菌活性   总被引:1,自引:0,他引:1       下载免费PDF全文
【背景】植物内生菌广泛分布在自然界中,具有巨大的潜在开发与应用价值。【目的】对温郁金内生细菌进行分离鉴定,并从中筛选出具有生防潜能的菌株。【方法】采用常规组织分离法对温郁金根茎内生细菌进行分离并利用16S rRNA基因序列分析进行初步鉴定;采用平板对峙法以铁皮石斛炭疽病菌(Colletotrichum gloeosporioides)为供试菌株对温郁金内生细菌进行拮抗菌的筛选;通过形态学鉴定、生理生化鉴定以及16S rRNA基因序列分析,确定拮抗菌株B-11的分类地位;以6个不同属的植物病原真菌为供试菌株对拮抗菌株B-11的抑菌谱进行测定,并研究其对病原菌菌丝的影响;利用特异性平板和MALDI-TOF-MS检测技术对拮抗菌株B-11产生的抑菌物质进行检测;采用离体叶片接种法研究拮抗菌株B-11对铁皮石斛炭疽病菌的防治效果。【结果】从温郁金根茎中分离得到25株内生细菌菌株,这些细菌菌株分属于12个属,其中芽孢杆菌属为优势菌属,占分离菌株的28%;通过初筛获得8株对C.gloeosporioides有抑菌活性的菌株,其中菌株B-11的抑菌活性最强,经鉴定该菌为贝莱斯芽孢杆菌(Baclliusvelezensis);抑菌谱测定发现菌株B-11能够对供试的属于不同属的6种植物病原真菌的菌丝生长具有抑制作用,显微观察发现经对峙培养6 d后这6种病原菌的菌丝体出现畸形膨大、分枝增多等现象;特异性平板检测结果表明拮抗菌株B-11能够产生蛋白酶、β-葡聚糖酶和嗜铁素,但不产生几丁质酶;MALDI-TOF-MS检测结果表明拮抗菌株B-11能够产生伊枯草素、丰源素和表面活性素3种脂肽类抗生素,其中伊枯草素的产量最高;离体叶片接种实验表明,拮抗菌B-11的离心去菌发酵液对铁皮石斛炭疽病的防治效率可达64%。【结论】温郁金根茎含有丰富的内生细菌资源,其内生细菌菌株B-11有潜力作为开发抗真菌代谢物和新药物的重要微生物资源。  相似文献   

8.
根据细菌核糖体基因16S rRNA保守端400 bp大小区段特征设计引物,应用聚合酶链式反应连接的限制性片段长度多态性(PCR-RFLP)方法建立一种新型灵敏的食源性致病菌鉴定技术。首先用特异性引物对9属19种细菌基因进行PCR扩增,扩增产物应用7种限制性内切酶Eco52Ⅰ、HindⅢ、MboⅠ、MluⅠ、PstⅠ、SalⅠ和XbaⅠ进行消化酶切,再利用琼脂糖凝胶电泳分辨酶切DNA片段大小数目,分析不同种属细菌酶切产物态型,从而进行基因型鉴别,针对16S rRNA基因片段利用RFLP进行分析,结果表明:19种致病菌表现出10种特异性的RFLP图谱,可准确鉴定区分9属细菌,最低检测限可以达到1 pg/μL。这证明PCR-RFLP技术可用于常见食源性致病菌的快速鉴定。  相似文献   

9.
Biolog细菌自动鉴定系统应用初探   总被引:19,自引:0,他引:19  
利用BiloogMicrostation细菌自动鉴定系统(3.50版)对已知的9个属23株菌进行了鉴定。24hBiolog系统鉴定结果:12林革兰氏阴性菌中,9株可准确鉴定到种的水平(75%),3株未达到属的水平。11株革兰氏阳性菌均属于芽孢杆菌属,全部鉴定到属的水平,9株准确鉴定到种的水平(81.8%)。总计属的水平准确率86.9%(20/23),种的水平准确率78.2%(18/23)。  相似文献   

10.
贵州半夏块茎腐烂病病原菌的分离与鉴定   总被引:2,自引:0,他引:2  
【目的】对贵州省半夏块茎腐烂病病原菌进行分离和鉴定,为开发有效的防治技术体系提供依据。【方法】采用组织分离法进行病原菌分离,根据柯氏法则进行致病性测定,结合形态学、生理生化特性和分子生物学方法进行鉴定。【结果】从贵州3个半夏产地患病样品中分离出12株病原细菌和5株病原真菌。经形态学、生理生化特性和分子生物学方法鉴定,12株病原细菌为胡萝卜软腐果胶杆菌胡萝卜软腐亚种;ZJ、Z3病原真菌为尖孢镰刀菌,D3、D5、H1为茄病镰刀菌。【结论】贵州省半夏块茎腐烂病的病原菌有病原细菌和病原真菌。  相似文献   

11.
MALDI-TOF-MS systems (Microflex-Bruker Daltonics/BioTyper? and Axima-Assurance-Shimadzu/SARAMIS-AnagnosTec) were assessed for bacterial identification. Focusing on bacteria difficult to identify routinely, 296 strains were identified by molecular biology techniques as gold standard. MALDI-TOF-MS identification provided correct results at genus and species level for 94.9%, 83.4% and 83.8%, 65.9% with Biotyper and Saramis respectively.  相似文献   

12.
太子参连作对根际土壤微生物的影响   总被引:2,自引:0,他引:2  
采用微生物培养法和末端限制性片断长度多态性(T-RFLP)技术分析连作对太子参根际土壤微生物多样性的影响。结果表明:连作导致太子参根际土壤细菌和好气性自生固氮菌数量极显著下降,相反,真菌、放线菌、厌气性纤维素分解菌数量极显著增加,而硝化细菌数量变化不显著。T-RFLP分析显示:与太子参-水稻-太子参轮作的土壤相比,太子参连作的土壤细菌种(属)略有减少,其中致病菌和病原菌种(属)增多,并出现一些具拮抗功能的链霉菌属(种);真菌种(属)则表现出上升的趋势,但未检索到与植物致病相关的真菌种(属)。  相似文献   

13.
The genus Saccharomyces comprises very closely related species. This high degree of relationship makes a simple identification and differentiation of strains difficult since these species are hardly discriminable by their morphological and physiological features. A sequence analysis of ribosomal DNA and the corresponding internal transcribed spacers can only rarely be successfully applied. In this study, we proved the applicability of a novel DNA fingerprinting method, the SAPD-PCR (specifically amplified polymorphic DNA) and of MALDI-TOF-MS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry) fingerprinting with the MALDI Biotyper for the differentiation of species belonging to the genus Saccharomyces. It was possible with SAPD-PCR to create specific banding patterns for all Saccharomyces species. Different strains of the same species produced nearly the same banding patterns. Specific and reproducible reference spectra could be generated for each of the strains with the MALDI Biotyper. Therefore, SAPD-PCR and MALDI-TOF-MS can be fast and reliable tools to identify these related Saccharomyces species which are applied in many biotechnological processes.  相似文献   

14.
The DNA-DNA hybridization method was used to study 4 species of bacteria of the genus Listeria. Concerning the DNA homology, L. monocytogenes strains may be divided into several species (in particular, the pathogenic forms may be isolated into independent taxon), in correlation with their biochemical and serological properties. Most of the studied strains of this species exhibit a high level of DNA homology--65-100%. Bacteria of the L. grayi and L. murrayi species are closely related to each other (90% of DNA homology), the reasonable suggestion being to unite them into a single species. L. denitrificans has 7% of DNA homology with the DNA of the other three species suggesting that it should be excluded from the genus Listeria.  相似文献   

15.

Background  

Heme is typically a major iron source for bacteria, but little is known about how bacteria of the Leptospira genus, composed of both saprophytic and pathogenic species, access heme.  相似文献   

16.
The relation between the number of some trinucleotides in the sequence of 16S rRNA gene and pathogenicity of bacterial species from the genera of Bacillus and Clostridium was revealed. The species of genus Bacillus, which are pathogenic for humans, mammals and insects, have an increased number of AAA and TAT triplets in 16S rRNA gene. Theoretically, these species, B. anthracis and B. cereus for example, may be detected in the specimen by the higher ratio of AAA plus TAT triplets to the number of GGG triplet. Species of genus Clostridium, which are pathogenic for humans and mammals, have a maximum ratio of AAA and TAT triplet numbers. This ratio was higher than 2.6 for pathogenic species and lower than 2.2 for saprophytic ones. These theoretical data may open a new way for detecting pathogenic bacteria through the determination of triplet numbers in the sequences of 16S rRNA or rRNA. However, the mechanism of evolutionary relation between the number of AAA and TAT triplets in the sequence of 16S rRNA gene and the pathogenicity of bacterial species is not known.  相似文献   

17.
Pyrazinamidase activity in 330 strains of bacteria from Enterobacteriaceae family (14 genus, 27 species) has been assessed. Pyrazinamidase activity detected in species from following genuses: Citrobacter, Escherichia, Klebsiella, Kluyvera, Morganella, Providencia, Raourtella, Salmonella, Shigella, and also in Proteus mirabilis, and nonpathogenic serovars of Yersinia enterocolitica, Y. frederiksenii. Pirasinamidase was absent in Serratia (S. marcescens, S. liguefaciens), Hafnia alvei, P. vulgaris, P. penneri, Y. pseudotuberculosis and pathogenic serovars of Y. enterocolitica. Absence of pyrazinamidase activity in bacteria from Hafnia and Serratia genus is a key taxonomic characteristic for identification of enterobacteria with microvolume assay technology.  相似文献   

18.
To identify the Yersinia genus and pathogenic species (Y. pestis, Y. pseudotuberculosis, Y. enterocolitica) in a single reaction a multiplex PCR technique was developed. It was optimized by five compounds of PCR buffer and temperature of primers annealing. The multiplex PCR provides an improved and rapid method for detection of the Yersinia genus and identification of pathogenic species.  相似文献   

19.
The Burkholderia genus consists of over 40 Gram-negative, beta-proteobacteria species that occupy remarkably diverse ecological niches. This genus contains species pathogenic to human, animals, and plants, as well as species involved in promoting plant growth and biodegradation of pollutants. This is largely explained by the extraordinary versatility of Burkholderia, as reflected by the remarkable diversity of extracellular products released by these bacteria. We exhaustively surveyed the extracellular enzymes, siderophores, toxins, antimicrobials, and other secondary metabolites produced by the members of this very diverse genus. Available information on regulation, especially quorum sensing mechanisms, and secretion is highlighted.  相似文献   

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