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1.
Biolog细菌自动鉴定系统应用初探 总被引:20,自引:0,他引:20
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
2.
真菌还原Cr(VI)的研究 总被引:7,自引:0,他引:7
从不同来源的样品中分离筛选出几株抗Cr(VI)的真菌,他们能在含300 ̄500mg/LK2Cr2O7的蔗糖合成培养基中生长,其中BS-1菌株抗K2Cr2O7达900mg/L.BS-1等4株真菌在含200mg/L K2Cr2O7的培养基中生长4 ̄6d后,培养液中的Cr(VI)已全部消失。这些真菌经鉴定为青霉菌BS-1和BS-3,黑曲霉BR-4和黄曲霉BX-1。经紫外可见光扫描及化学分析证实,高毒的C 相似文献
3.
以 Cytopore 多孔微球固定产重组组织型纤溶酶原激活剂(rt P A) C H O 工程细胞株4 B3 ,在2 L 搅拌式生物反应器用无血清培养基 D F5 S 连续灌流培养。4 B3 细胞的最大活细胞密度和rt P A 生产水平分别达到883 ×106/ m L 和12473 I U/ m L。含rt P A 的4 B3 细胞培养上清经 M P G 吸附层析和 Lysinesepharose 4 B 亲和层析两步纯化,rt P A 的纯度达到98 % 。 相似文献
4.
从土壤中分离到一株黄杆菌(Flavobacteriumspp)CF-60,该菌的生长需Mg2+存在,MgSO4·7H2O的最适浓度为0.2%;蛋白胨是该菌株生长的最好氮源,它不能利用无机氮。种龄超过96h的菌体不能在新鲜培养基中生长。经54h的2L恒化器发酵,生物量达6.8g/L,色素产量为10.6mg/L。该菌产生的类胡萝卜素成分简单,主要成分的含量为90.3%,该成分经初步鉴定是分子结构中含有羰基和羟 相似文献
5.
从不同来源的细菌菌株筛选获得一株吸附还原Au3+较强的菌株D01,经鉴定为巨大芽孢杆菌(Bacilusmegatherium)D01。菌株D01在Au3+浓度600mg/L下仍能较好生长。从电化学反应表明,该菌具有较强的还原力,它能将金催化剂的前驱体Au3+/αFe2O3还原成具有催化CO+O2→CO2的高分散度的Au0/αFe2O3催化剂 相似文献
6.
BIOLOG系统鉴定黄瓜根围促生菌的初步研究 总被引:8,自引:0,他引:8
对筛选出的8株具有明显促生防病作用的黄瓜根围促生菌CN11,CN31,CN45,CN1 16,CN129,XB120,XB5,XB41通过BIOLOG法进行了分类鉴定,分别为:铜绿假单胞菌(Psendomonas aeruginosa),波纹假单胞菌(P. Corrugata),短芽孢杆菌(Bacillus brevis),荧光假单胞菌B型(P. Fluorescens type B),铜绿假单胞菌(P. Aeruginosa),荧光假 相似文献
7.
黑山晋枣芽培养及值株再生研究 总被引:4,自引:0,他引:4
以珍稀、优质黑山晋枣液芽为外植体建立了原种无性繁殖系。继代繁殖培养基(mg/L):MS+BA0.1+IAA0.1和MS+BA0.1+IBA0.5,培养30d平均苗高3.0-3.3cm,芽繁殖系数3.3-3.5;生根培养基为MS+IAA0.1+IBA0.2及MS+IAA0.2+IBA0.2两种,生根率83.3%,培养40d平均具根苗高达4.99cm,一次成苗,简化程序,缩短成苗期。移载成活率83.0 相似文献
8.
在 B. Braun E S10 型15 L 和 N B S Bio Flo 3000 型5 L 发酵罐中,利用补料分批培养技术高密度表达培养含重组质粒p S B H L11 的大肠杆菌 Y K537 ,生产重组人白细胞介素3( I L3) ,发现在发酵过程中,限制性流加甘油,控制溶解氧在30 % ~40 % 左右、30 ℃生长11h ,42 ℃诱导培养4h ,能将发酵液中最终菌体密度从 O D16600 提高到 O D53600( 相当于每升发酵液含106 克湿菌体) ,并且保持了白细胞介素3 的表达量,占菌体总蛋白的30 % 左右,含量超过33 % g/ L,使 I L3 包涵体产量从湿重22g/ L 提高到85 g/ L,纯化步骤比较简单,超声破菌后经两次洗涤纯度就达到70 % 以上。 相似文献
9.
芽孢杆菌M_(50)产生β甘露聚糖酶的条件研究 总被引:3,自引:0,他引:3
从土壤中分离到9 株产生β甘露聚糖酶的芽孢杆菌( Bacillus sp .) 。Bacillussp . M50250m L三角瓶摇瓶培养试验,以4 % 的魔芋粉为碳源,1-0 % ( NH4)2SO4 为氮源,0-35 %Na2CO3 ,30 ~34 ℃培养60h 产酶达到高峰。酶活力为180 ~200u/m L。100L 罐发酵,在30 ~32 ℃,1∶0 .75vvm 通气量,200r/min 条件下,发酵液酶活力高达330u/m L。酶的最适反应温度和pH 分别为50 ℃和6-0 ,低于50 ℃,pH5 .0 ~7 .0 酶稳定。Fe3+ 、Al3+ 、EDTA、Hg2+ 对酶有抑制作用,而Ba2+ 、Mn2+ 对酶有激活作用。发酵粗酶液对苎麻精干麻精练,显示对精干麻的半纤维素残胶具有降解作用。 相似文献
10.
给DBA/2小鼠双前肢足垫皮下注射2×10P388细胞后,腹腔注入L-IFNr(脂质体包裹IFNr)或IFNr4,20,100×104U/(kg·d),连续10d,结果小鼠生存期分别延长18.3、29.3、33.9、23.9、31;9、33.6d。在第15天循环血中的单核细胞分别相应降低25.3%、54.9%,89.7%和55.8%、85.2%、90.2%;移植瘤引流区腋窝淋巴结肿大呈剂量依赖性缩小。结果表明两种IFNr制剂均有显著的抗白血病细胞P388转移的作用,L-IFNr的效果优于IFNr。 相似文献
11.
Klingler JM Stowe RP Obenhuber DC Groves TO Mishra SK Pierson DL 《Applied and environmental microbiology》1992,58(6):2089-2092
Biolog's identification system was used to identify 39 American Type Culture Collection reference taxa and 45 gram-negative isolates from water samples. Of the reference strains, 98% were identified to genus level and 76% to species level within 4 to 24 h. Identification of some authentic strains of Enterobacter, Klebsiella, and Serratia was unreliable. A total of 93% of the water isolates were identified. 相似文献
12.
J M Klingler R P Stowe D C Obenhuber T O Groves S K Mishra D L Pierson 《Applied microbiology》1992,58(6):2089-2092
Biolog's identification system was used to identify 39 American Type Culture Collection reference taxa and 45 gram-negative isolates from water samples. Of the reference strains, 98% were identified to genus level and 76% to species level within 4 to 24 h. Identification of some authentic strains of Enterobacter, Klebsiella, and Serratia was unreliable. A total of 93% of the water isolates were identified. 相似文献
13.
Simon Lévesque Philippe J. Dufresne Hafid Soualhine Marc-Christian Domingo Sadjia Bekal Brigitte Lefebvre Cécile Tremblay 《PloS one》2015,10(12)
Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has emerged as a rapid, highly accurate, and cost-effective method for routine identification of a wide range of microorganisms. We carried out a side by side comparative evaluation of the performance of Bruker Biotyper versus VITEK MS for identification of a large and diverse collection of microorganisms. Most difficult and/or unusual microorganisms, as well as commonly encountered microorganisms were selected, including Gram-positive and negative bacteria, mycobacteria, actinomycetes, yeasts and filamentous fungi. Six hundred forty two strains representing 159 genera and 441 species from clinical specimens previously identified at the Laboratoire de santé publique du Québec (LSPQ) by reference methods were retrospectively chosen for the study. They included 254 Gram-positive bacteria, 167 Gram-negative bacteria, 109 mycobacteria and aerobic actinomycetes and 112 yeasts and moulds. MALDI-TOF MS analyses were performed on both systems according to the manufacturer’s instructions. Of the 642 strains tested, the name of the genus and / or species of 572 strains were referenced in the Bruker database while 406 were present in the VITEK MS IVD database. The Biotyper correctly identified 494 (86.4%) of the strains, while the VITEK MS correctly identified 362 (92.3%) of the strains (excluding 14 mycobacteria that were not tested). Of the 70 strains not present in the Bruker database at the species level, the Biotyper correctly identified 10 (14.3%) to the genus level and 2 (2.9%) to the complex/group level. For 52 (74.2%) strains, we obtained no identification, and an incorrect identification was given for 6 (8.6%) strains. Of the 178 strains not present in the VITEK MS IVD database at the species level (excluding 71 untested mycobacteria and actinomycetes), the VITEK MS correctly identified 12 (6.8%) of the strains each to the genus and to the complex/group level. For 97 (54.5%) strains, no identification was given and for 69 (38.7%) strains, an incorrect identification was obtained. Our study demonstrates that both systems gave a high level (above 85%) of correct identification for a wide range of microorganisms. However, VITEK MS gave more misidentification when the microorganism analysed was not present in the database, compared to Bruker Biotyper. This should be taken into account when this technology is used alone for microorganism identification in a public health laboratory, where isolates received are often difficult to identify and/or unusual microorganisms. 相似文献
14.
A.M. Day J.A.T. Sandoe J.H. Cove & M.K. Phillips-Jones 《Letters in applied microbiology》2001,33(5):392-396
AIMS: To evaluate the full test scheme of Facklam and Sahm (1995) for the identification of clinical enterococcal isolates to genus and species level. METHODS AND RESULTS: Fifty-nine clinical isolates, previously provisionally classed as enterococci on the basis of just four biochemical tests of Facklam and Sahm and one other test, were subjected to genus and species identification using the full identification scheme of Facklam and Sahm; 98% of these strains were confirmed to be enterococci and of these, 69% were identified as Enterococcus faecalis and 31% as Enterococcus faecium. Six tests in the scheme (out of 24) gave anomalous or unreliable results for some strains, and two gave unexpected results for the majority of strains presumptively identified as Ent. faecium. CONCLUSIONS: Nine (out of 12) genus tests and nine (out of 12) species tests from the Facklam and Sahm scheme were reliable. Testing for the presence of the Lancefield antigen D was also useful. The majority of presumptive Ent. faecium strains gave different results for the sorbitol and raffinose tests from that expected. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates the level of reliability for each of the tests in a current enterococcal identification scheme for differentiating clinical isolates, and showed that two tests gave consistently different test results from those expected for Ent. faecium. 相似文献
15.
Accurate identification of bacterial isolates is an essential task in clinical microbiology. Phenotypic methods are time-consuming and either fail to identify some bacteria such as Gram-positive rods entirely or at least fail to do so in some clinical situations. 16S rDNA sequencing is a recent method of identification which offers a useful alternative. In this study, we investigate the usefulness of this method for identifying a range of bacteria in a clinical laboratory under routine conditions. Over a period of 30 months, 683 isolates were obtained from clinical specimens, sequenced and analysed. For 568 of these isolates (83.1%), the sequence provided species level identification. For 108 isolates (15.8%), the identification was limited to the genus level, and for 7 isolates (1%), the sequence remained unidentifiable by 16S rDNA sequence analysis. For the isolates identified only to the genus level, the 16S rDNA approach failed to identify bacteria to the taxonomic level for 3 reasons: failure to differentiate between species in 72 isolates (66%), the lack of any closely related sequence in the database for 15 isolates (13.8%) and the presence of more than 1% of undetermined position in the sequence for 13 isolates (12%). 相似文献
16.
Moore DF Zhowandai MH Ferguson DM McGee C Mott JB Stewart JC 《Journal of applied microbiology》2006,100(6):1272-1281
AIMS: To compare accuracy of genus and species level identification of presumptive enterococci isolates from the marine environment using conventional biochemical testing, four commercial identification systems and 16S rRNA sequence analysis. METHODS AND RESULTS: Ninety-seven environmental bacterial isolates identified as presumptive enterococci on mEI media were tested using conventional and Enterococcus genus screen biochemical tests, four commercial testing systems and 16S rRNA sequencing. Conventional and Enterococcus genus screen biochemical testing, 16S rRNA sequencing and two commercial test systems achieved an accuracy of > or = 94% for Enterococcus genus confirmation. Conventional biochemical testing and 16S rRNA sequencing achieved an accuracy of > or = 90% for species level identification. CONCLUSIONS: For confirmation of Enterococcus genus from mEI media, conventional or genus screen biochemical testing, 16S rRNA sequencing and the four commercial systems were correct 79-100% of the time. For speciation to an accuracy of 90% or better, either conventional biochemical testing or 16S rRNA sequencing is required. SIGNIFICANCE AND IMPACT OF THE STUDY: Accurate identification of presumptive environmental Enterococcus isolates to genus and species level is an integral part of laboratory quality assurance and further characterization of Enterococcus species from pollution incidents. This investigation determines the ability of six different methods to correctly identify environmental isolates. 相似文献
17.
Cirković I Hauschild T Jezek P Dimitrijević V Vuković D Stepanović S 《Current microbiology》2008,57(2):158-160
This study evaluated the performance of the BD Phoenix system for the identification (ID) and antimicrobial susceptibility testing (AST) of Staphylococcus vitulinus. Of the 10 S. vitulinus isolates included in the study, 2 were obtained from the Czech Collection of Microorganisms, 5 from the environment, 2 from human clinical samples, and 1 from an animal source. The results of conventional biochemical and molecular tests were used for the reference method for ID, while antimicrobial susceptibility testing performed in accordance with Clinical and Laboratory Standards Institute recommendations and PCR for the mecA gene were the reference for AST. Three isolates were incorrectly identified by the BD Phoenix system; one of these was incorrectly identified to the genus level, and two to the species level. The results of AST by the BD Phoenix system were in agreement with those by the reference method used. While the results of susceptibility testing compared favorably, the 70% accuracy of the Phoenix system for identification of this unusual staphylococcal species was not fully satisfactory. 相似文献
18.
Shunsuke Segawa Motoi Nishimura Kazuyuki Sogawa Sachio Tsuchida Shota Murata Masaharu Watanabe Kazuyuki Matsushita Katsuhiko Kamei Fumio Nomura 《Clinical proteomics》2015,12(1)
Background
The MALDI (matrix-assisted laser desorption/ionization) Biotyper system for bacterial identification has already been utilized in clinical microbiology laboratories as a successful clinical application of protoemics. However, in cases of Nocardia, mass spectra suitable for MALDI Biotyper identification are often not obtained if such specimens are processed like general bacteria. This problem is related to the insufficiencies in bacterial spectrum databases that preclude accurate specimen identification. Here, we developed a bacterial processing method to improve mass spectra from specimens of the genus Nocardia. In addition, with the new processing method, we constructed a novel in-house bacterial database that combines a commercial database and mass spectra of Nocardia strains from the Department of Clinical Laboratory at Chiba University Hospital (DCLC) and the Medical Mycology Research Center at Chiba University (MMRC).Results
The newly developed method (Nocardia Extraction Method at DCLC [NECLC]) based on ethanol-formic acid extraction (EFAE) improved mass spectra obtained from Nocardia specimens. The Nocardia in-house database at Chiba University Hospital (NDCUH) was then successfully validated. In brief, prior to introduction of the NECLC and NDCUH, 10 of 64 (15.6%) clinical isolates were identified at the species level and 16 isolates (25.0%) could only be identified at the genus level. In contrast, after the introduction, 58 isolates (90.6%) were identified at the species level and 6 isolates (9.4%) were identified at the genus level.Conclusions
The results of this study suggest that MALDI-TOF (time-of-flight) Biotyper system can identify Nocardia accurately in a short time in combination with a simple processing method and an in-house database.Electronic supplementary material
The online version of this article (doi:10.1186/s12014-015-9078-5) contains supplementary material, which is available to authorized users. 相似文献19.
Sameer A. Barghouthi 《Indian journal of microbiology》2011,51(4):430-444
The Universal Method (UM) described here will allow the detection of any bacterial rDNA leading to the identification of that
bacterium. The method should allow prompt and accurate identification of bacteria. The principle of the method is simple;
when a pure PCR product of the 16S gene is obtained, sequenced, and aligned against bacterial DNA data base, then the bacterium
can be identified. Confirmation of identity may follow. In this work, several general 16S primers were designed, mixed and
applied successfully against 101 different bacterial isolates. One mixture, the Golden mixture7 (G7) detected all tested isolates
(67/67). Other golden mixtures; G11, G10, G12, and G5 were useful as well. The overall sensitivity of the UM was 100% since
all 101 isolates were detected yielding intended PCR amplicons. A selected PCR band from each of 40 isolates was sequenced
and the bacterium identified to species or genus level using BLAST. The results of the UM were consistent with bacterial identities
as validated with other identification methods; cultural, API 20E, API 20NE, or genera and species specific PCR primers. Bacteria
identified in the study, covered 34 species distributed among 24 genera. The UM should allow the identification of species,
genus, novel species or genera, variations within species, and detection of bacterial DNA in otherwise sterile samples such
as blood, cerebrospinal fluid, manufactured products, medical supplies, cosmetics, and other samples. Applicability of the
method to identifying members of bacterial communities is discussed. The approach itself can be applied to other taxa such
as protists and nematodes. 相似文献