细菌鉴定仪出现“不能鉴定细菌”的处理

我们对现今应用较为普遍的ＶＩＴＥＫ ＡＭＳ鉴定系统中报告“不能鉴定细菌”(ＵＩＯ)菌株作了进一步分析 ,探讨其发生原因、处理方法 ,以期建立一套切实可行的仪器与手工方法相结合的细菌鉴定方法。一、材料与方法1 仪器与试剂 :ＶＩＴＥＫ ＡＭＳ 6 0型全自动细菌鉴定仪及其配套ＧＮＩ、ＧＰＩ、ＹＢＣ试验卡 (生物梅里埃公司产品 )。2 试验菌株 :10 2 5株本院 1999.10 .2 1 2 0 0 0 .3.3临床标本分离的菌株进行了Ｖｉｔｅｋ ＡＭＳ鉴定 ,其中应用ＧＮＩ卡 6 31例、ＧＰＩ卡 334例、ＹＢＣ卡 6 0例。3 ＶＩＴＥＫ ＡＭＳ鉴定法 :根据细菌…     

利用细菌鉴定卡保存菌种

微生物实验室成功地进行细菌菌种保存 ,对于系统地分析流行菌株的生物学、分子生物学特征和耐药性的变迁 ,以及对医院感染病原学调查、新药临床研究都十分重要[1] 。我们通过对 1998年 1月～ 2 0 0 0年 5月两年多时间冰冻保存的ＶＩＴＥＫ ＧＮＩ、ＧＰＩ、ＹＢＣ 3种生化鉴定卡共 470 0株常见的细菌 ,选择其中 2 5 8株常见进行复活再鉴定 ,以探讨一种操作简便而又实用可靠的保存临床标本分离细菌菌株的方法 ,现报道如下。一、材料和方法1 标本来源 :2 5 8株 1998年 1月～ 2 0 0 0年 5月从痰、血、尿、脓液等标本中分离、纯培养的细菌 ,用Ｖ…     

临床分离肠球菌耐药性和耐药基因分布及其相关性研究

目的:研究临床分离肠球菌的耐药性、耐药基因分布及其相关性.方法:采用Vitek2 compact全自动细菌鉴定及药敏分析系统进行临床分离肠球菌的鉴定和药敏试验;提取细菌基因组DNA,通过PCR法检测14种肠球菌耐药相关基因;采用SPSS25.0软件对肠球菌耐药性和耐药基因相关性进行统计学分析.结果:临床分离的89株肠球...     

鲍曼不动杆菌喹诺酮耐药决定区基因突变的研究

随着喹诺酮类药物的广泛应用 ,鲍曼不动杆菌对其敏感性逐渐降低[1] 。本研究对临床分离的鲍曼不动杆菌喹诺酮耐药决定区的ｇｙｒＡ和ｐａｒＣ基因突变情况进行了分析 ,报告如下。一、材料和方法1.细菌鉴定 :从临床分离菌中经ＶＩＴＥＫ细菌鉴定系统鉴定为鲍曼不动杆菌 2 3株。2 .药敏试验 :采用浓度梯度法 (Ｅｔｅｓｔ)试条测定其对环丙沙星最低抑菌浓度 (ＭＩＣ)值。3 .菌株分类 :按美国临床实验室标准化委员会 2 0 0 0年版标准分类 :12株为耐药菌 ,其中ＭＩＣ≥ 32 μｇ/ｍｌ9株 ,4～ 16μｇ/ｍｌ 3株 ;11株为敏感菌 ,ＭＩＣ值为 0 2 5～…     

慢性鼻窦炎患者窦道脓液的细菌培养结果分析

细菌感染是引起慢性鼻窦炎的病因之一。为了解本病患者中鼻道、上颌窦及筛窦 3个部位的细菌分布 ,我们选择了 2 0 5例慢性鼻窦炎患者脓液标本做细菌培养 ,并筛选具有临床意义的细菌进行分离鉴定及药敏试验。一、材料与方法1.标本来源 :标本的采集是按照美国ＡＳＭ《临床微生物学手册》中标本采集送检指南操作 ,均为合格标本。选择2 0 0 1年 4～ 8月确诊为慢性鼻窦炎的住院患者 ,在功能性内窥镜手术中 ,用特制的棉签 ,经严格消毒后 ,无菌采集 3个部位脓液标本各 1份于厌氧运送培养基中 ,立即送检 ,分别作普通和厌氧培养。2 .抗生素纸片及质控…     

临床分离细菌的菌种分布及耐药分析

材料与方法 1.菌株来源:菌株取自2002～2003年住院患者送检的各类标本,共分离病原菌357株. 2.细菌鉴定及药敏试验:采用法国生物梅里埃公司鉴定条进行细菌鉴定;药敏试验采用纸片扩散法.操作及结果判读均按1999年NCCLS规定进行,全部培养基购自生物梅里埃公司,药敏纸片购自北京天坛生物技术有限公司.     

血培养阳性标本直接细菌鉴定和药敏中应用分离胶试管的方法探讨

目的探讨血培养的细菌快速鉴定和药敏实验,与常规法(转种后取得的纯菌落鉴定及药敏试验)的符合率。方法血培养仪器报警阳性后,无菌吸取一定量的培养液至含分离胶的试管中,2 000 rpm离心15 min后吸取分离胶上层含菌液体,直接进行细菌鉴定和药物敏感试验;同时与转种后取得的纯菌落鉴定及药敏试验常规方法结果进行比较。结果 66株血培养阳性标本离心集菌直接鉴定和转种菌落常规鉴定结果符合率达85%以上;药敏试验结果符合率>95%。离心集菌法比常规法提前近24 h。结论血培养阳性标本使用分离胶的试管离心集菌后直接进行细菌鉴定及药敏试验,可以缩短检验时间,且与常规法的实验结果符合率高。     

产超广谱β-内酰胺酶细菌的检测及其药敏试验结果分析

由于 β 内酰胺酶类抗生素的广泛应用及不合理使用 ,耐药菌株迅速增加 ,尤其产超广谱 β 内酰胺酶(ESBLs)细菌引起的耐药问题更为严重。为了解本院ESBLs细菌的检出及耐药情况 ,我们对从住院患者的细菌送检标本中分离得到的 15 2株大肠埃希菌、肺炎克雷伯菌进行了ESBLs检测 ,并对其进行药敏试验 ,结果报告如下。1　材料与方法1.1　菌株来源　 2 0 0 1年 7月至 2 0 0 2年 6月从本院住院患者临床细菌标本中分离的菌株 ,其中大肠埃希菌 10 6株 ,肺炎克雷伯菌 4 6株。分离的菌株在半固体琼脂中保存。1.2　菌株鉴定　用ATB细菌分析系统 (法…     

临床细菌检验快速方法应用进展

临床细菌检验程序包括标本分离、菌种鉴定及药敏试验３个步骤，其中菌种鉴定的快慢是影响整个检验时间的关键。因此临床细菌检验快速方法的研究也多以菌种鉴定为主。一、用分离培养基对菌种直接鉴定在分离培养基中加入检测某些菌种特异性酶的底物，在菌种分离的同时直接进...     

重症监护病房鲍曼不动杆菌21株抗菌药物敏感性分析

目的 :了解重症监护病房 (ICU)鲍曼不动杆菌肺炎药物敏感性特点 ,为临床防治提供指导。方法 :鲍曼不动杆菌的分离和鉴定用ATB细菌鉴定仪 ,以琼脂稀释法对分离菌株进行药物敏感性测定。结果 :对于本次试验的 2 1株鲍曼不动杆菌 ,亚胺培南的抗菌活性最强 ,敏感率为 90 % ,其次是头孢哌酮 舒巴坦 ,敏感率为 81% ;阿米卡星、头孢吡肟、头孢他啶的敏感率在6 0 %左右 ;细菌对于头孢唑林及氨苄西林的敏感率 <5 %。结论 :ICU鲍曼不动杆菌肺炎经验治疗可选用亚胺培南、头孢哌酮 舒巴坦。合理应用抗生素及做好消毒隔离 ,规范操作等措施可有效防治ICU鲍曼不动杆菌肺炎     

A enzymatic and fluorimetric determination of total bile acid sulfates in human urine

A simple, precise and sensitive method for separation and determination of total bile acid sulfates in human urine is described. The sulfate fraction of urinary bile acids was separated with lipophilic anion exchange gel, piperidinohydroxypropyl Sephadex LH-20 after sample clean-up with Sep-Pak C18 cartridge. The obtained sulfate fraction was submitted to solvolysis with a small volume of dimethoxypropane-HCl solution and subjected to enzymatic-fluorimetrical assay using 3 alpha-hydroxysteroid dehydrogenase and resazurin. In this method, no influence of existing salts in the reaction mixture on fluorescence intensity was observed and solvolysis reaction was almost complete. Overall recoveries of glycine- and taurine-conjugated bile acid 3-sulfates from normal urine ranged from 90.5 to 93.7% and those of unconjugates from 48.7 to 78.0%. The sensitivity of the described method enabled to estimate total bile acid sulfates with 0.5 ml of normal urine and precision tests showed the satisfactory accuracy. The level of total urinary bile acid sulfates was estimated on some patients with hepatobiliary diseases and healthy subjects.     

Clinical chemistry of 3-methoxy-17-epioestriol (epimestrol)

The possibilities of clinical chemical analysis of 3-methoxy-17-epioestriol (Epimestrol) were investigated. Infrared and visible absorption curves were recorded and the possibility of its determinations shown. The method of ether extraction from aqueous medium is described. Separation from oestrogens was investigated by column and thin-lay er chromat ography.A new gas-chromatographic method of separation and determination is described, by means of which 2 μg Epimestrol can be determined.     

Improved method for investigation of copper metabolism in patients with Wilson's disease using Cu

A radiochemical method for the separation of albumin- and ceruloplasminbound ⁶⁴Cu from serum is described. Separation was achieved by column chromatography with an iminodiacetic acid-chelating resin at pH 5.0 and 40°. The experimental details for the separation are given.     

Sterile isolation of polymorphonuclear leukocytes from large blood volumes

A method for preparation of pure polymorphonuclear leukocyte populations from large blood volumes is described. Methyl cellulose sedimentation and hypotonic haemolysis resulted in an efficient removal of erythrocytes. Subsequent separation with sodium metrizoate/Ficoll (density 1.077) gave rise to a preparation containing 95% polymorphonuclear leukocytes with a recovery of 56% and complete exclusion of red cell ghosts. The granulocyte function expressed by chemotaxis to the standard attractant casein and by trypan blue exclusion was not influenced by the separation procedure, which may be applied prior to granulocyte labeling for studies of cell kinetics and inflammatory disease.     

免疫磁珠捕获法检测志贺菌的模拟研究

目的评估所制备的志贺菌免疫磁珠特异性富集作用,为建立基于免疫磁珠捕获法的志贺菌检测流程提供技术依据。方法制备志贺菌属细菌(鲍氏志贺菌除外)、非目标菌(大肠埃希菌、产气肠杆菌、鲍曼不动杆菌、肺炎克雷伯菌、弗尼斯弧菌、表皮葡萄球菌)的菌悬液,利用志贺菌特异性多抗免疫磁珠进行捕获,接种平板后进行菌落计数,并计算免疫磁珠的特异性及非特异性吸附效率。设计干扰试验,在含有104 cfu/mL的大肠埃希菌悬液中,加入不同浓度的目标菌,用所制备的磁珠进行捕获,评估免疫磁珠的特异性富集效率。在经3和6 h预增菌的含有已知量目标菌的奶粉模拟标本及未增菌的粪便模拟标本中,分别加入所制备的磁珠进行捕获,评估基于免疫磁珠捕获法的志贺菌检测流程的可行性。结果菌落计数表明,制备的志贺菌免疫磁珠对痢疾志贺菌、福氏志贺菌和宋内志贺菌的捕获效率均超过30%,对6种非目标菌的非特异性吸附效率<3%。干扰试验结果显示,与常规直接平板接种法相比,免疫磁珠捕获法的敏感性提高了约100倍;奶粉和粪便模拟试验的结果显示,免疫磁珠捕获法的最低检测限分别可达101 cfu/25 g和101 cfu/mL,并对标本中目标菌的检出率有一定提高。结论志贺菌免疫磁珠对志贺菌属细菌(鲍氏志贺菌除外)具有一定特异捕获与富集作用,可应用于实验室的日常检测工作。     

The measurement of serum free insulin by steady-state gel filtration

A method for the separation of bound and free insulin in the serum of insulin treated diabetics by steady state gel filtration is described. The method meets the criteria which have been suggested for the validation of a method of separating bound from free ligand. The precision of the method is satisfactory, and the results compare well with those obtained by the methods of ethanol precipitation and polyethylene glycol precipitation.     

Chromatographic separation of hemoglobins A 2 and C. The quantities of hemoglobin A 2 in patients with AC trait, CC disease, and C- -thalassemia

A method is described which allows the separation of Hb-A₂ and Hb-C in red cell hemolysates of subjects heterozygous or homozygous for this variant. The separation is obtained on columns of carboxymethyl cellulose using 0.01 M sodium phosphate buffers as developers. The levels of Hb-A₂ in Hb-C heterozygotes and homozygotes tend to be higher than those found in normal adults and are comparable to percentages reported for patients with sickle cell trait and sickle cell anemia.     

The estimation of true urinary tyramine in man

A method involving solvent extractions and chromatographic separation is described which permits the measurement of true free and conjugated tyramine in urine.     

A new screening test for the diagnosis of mucopolysaccharidoses.

A simple method for the electrophoretic separation of glycosaminoglycans from untreated urine is presented. It is shown that this rapid electrophoresis on specially shaped acetate cellulose strips gives more information than other screening tests used in laboratory for discriminating between normal and pathological samples. This technique is useful particularly with low levels of glycosaminoglycans which may give false-negative responses.     

Fluorometric determination of the oral hypoglycemic agent β-phenethylbiguanide using alkaline ninhydrin

A fluorometric method for determining β-phenethylbiguanide (PEBG) in microgram and submicrogram quantities is described. This method is based upon a reaction with alkaline ninhydrin. The lower practical limit of PEBG which can be detected by this method is 0.025 μg/ml. The method is not specific for PEBG and applicability for measurements in biological media will require separation of PEBG from other guanidinium compounds.