用于嗜血杆菌鉴定的卫星试验影响因素研究

目的了解嗜血杆菌鉴定中X、V因子测定的各种影响因素及可用于嗜血杆菌鉴定的卫星试验的细菌种类.方法用4种不同的琼脂平板,比较不同培养时间和CO2环境对嗜血杆菌X、V因子需求试验结果的影响,同时比较不同菌种在各种琼脂培养基上对嗜血杆菌琼脂卫星试验结果的影响.结果 X、V因子需求试验结果表明,脑心琼脂效果最好,其次为营养琼脂,M-H琼脂结果与脑心琼脂相比差异有显著性(P<0.01);CO2对试验结果有影响;培养48 h较24 h的菌落大,易于辨认;不同菌种在不同琼脂培养基上对嗜血杆菌琼脂卫星试验的结果有所不同.结论卫星试验应在CO2环境中进行,用脑心琼脂平板测试嗜血杆菌对X、V因子的需求结果较好,多数葡萄球菌及部分革兰阴性杆菌对嗜血杆菌的琼脂卫星试验结果较好.     

嗜血杆菌鉴定中X、V因子及溶血性测定影响因素探讨

目的：了解Haemophilus(嗜血杆菌）鉴定中X、V因子测定的各种影响因素。方法：用5种不同的琼脂平板，比较不同培养时间和8％CO2环境对嗜血杆菌X，V因子需求试验及溶血试验结果的影响。结果：X、B因子需求试验结果表明，脑心琼脂效果最好，其次为营养琼脂，MH琼脂与脑心琼脂结果比较有显著差异（P＜0．01），CO2对试验结果有影响；培养48h较24h的菌落大易辨认。结论：用脑心琼脂平板测试嗜血杆 对X、V因子的需求及以CO2环境中检测嗜血杆菌的溶血性结果较好。     

临床微生物实验室需重视嗜血杆菌的分离培养

嗜血杆菌属属巴斯德菌科,因在生长过程中需含有X因子和V因子的血液琼脂而得名[1]。人类感染常见菌种为流感嗜血杆菌、副流感嗜血杆菌、杜克雷嗜血杆菌,本属细菌主要寄生在人的咽喉及口腔黏膜,少见于消化道和生殖道[1-2]。其中流感嗜血杆菌主要引起人类的急性化脓感染,如急性咽炎、喉炎、气管炎、中耳炎、鼻窦炎、肺炎、心内膜炎、败血症等以及严重的继发感染。近年来儿童呼吸道感染嗜血杆菌呈上升趋势[3]。杜克雷嗜血杆菌是引起软下疳的病原菌。嗜血杆菌属苛氧菌,对其分离培养是反映实验室鉴定水平的标志[4]。许多     

嗜血杆菌在儿童及老年人上呼吸道中的分布调查

Haemophilus(嗜血杆菌)是苛养性细菌,与许多严重感染性疾病有关[1],其生长不仅需要X、V因子,而且受革兰阳性细菌生长的抑制,分离培养极为困难.我们用含50 mg/ L万古霉素的兔血巧克力琼脂培养基对121名60～97岁老年人及165名3～6岁儿童的咽部分泌物进行了嗜血杆菌分离和鉴定,并对所分离的嗜血杆菌进行了β-内酰胺酶测试,结果报告如下.     

探讨全自动快速微生物检测系统在鉴定流感嗜血杆菌的应用

目的探讨全自动快速微生物检测系统(VITEK MS)在鉴定流感嗜血杆菌的应用价值。方法收集广州市黄埔区红十字会医院与广东省中医院2014年3月至2015年1月从痰液或咽拭子标本中分离的流感嗜血杆菌35株,用16SrDNA测序确证。并且用VITEK MS质谱仪,API NH常规生化鉴定与MH琼脂平板上V+X因子需求性试验3种方法对上述菌株鉴定分析。结果在35株流感嗜血杆菌细菌中,VITEK MS质谱仪与API NH常规生化鉴定差异无统计学意义(P0.05),而VITEK MS与MH琼脂平板上V+X因子需求性试验差异有统计学意义(P0.05)。结论对于API NH常规生化鉴定与MH琼脂平板上V+X因子需求性试验,VITEK MS能快速、准确地鉴定流感嗜血杆菌,对临床诊治提供很大的帮助,有非常好的应用前景。     

一种新型嗜血杆菌分离培养基的评价与应用

本研究对一种新型的嗜血杆菌分离培养基与其他 4种常用嗜血杆菌培养基进行了对照 ,并统计了临床标本分离率 ,报告如下。1.标本来源 :痰、咽拭子、分泌物共 42 8份 ,来自我院。2 .试剂 :脑心浸液来自生物梅里埃公司及BBL公司 ,哥伦比亚琼脂来自DIFCO公司 ,酵母浸液、牛肉浸液为本室自制 ,脱纤维羊血、兔血来自我院动物室。3 .标准菌株 :流感嗜血杆菌ATCC492 47来自卫生部临床检验中心。4.培养基 :( 1)Schoc :营养琼脂 + 6%羊血 ;85℃水浴 10min ,凉后倾制平皿 ;( 2 )BBrchoc :BBL脑心浸液 +琼脂 + 6%兔血 ;方法同上 ;( 3 )MBrchoc :…     

无水乙醇对变形杆菌迁徙性生长的抑制效果评价

目的创建一种简便易行的抑制变形杆菌迁徙性生长的方法。方法用无水乙醇预处理营养琼脂平板(NUA)、哥伦比亚琼脂平板(CAN)和中国蓝平板(CBA),在平板中心点种奇异变形杆菌标准菌株,观察对该菌迁徙生长的抑制效果;分别划线接种4种标准菌种,培养24 h后进行鉴定和药敏试验。对照组:在未经乙醇处理的培养基中心点种奇异变形杆菌标准菌株。结果奇异变形杆菌于乙醇处理培养基37℃培养12、24、48 h均未出现迁徙生长现象。接种的4种标准菌株鉴定和药敏结果未发生改变。结论无水乙醇能有效地抑制变形杆菌的迁徙性生长现象,并且对常见的革兰阳性球菌、革兰阴性杆菌的生长特性和药敏结果无影响。     

732株嗜血杆菌对阿奇霉素敏感性分析

1999年 3月至 2 0 0 0年 12月 ,对南京军区南京总医院住院病人 ,门诊病人送检的痰液及咽拭标本中分离的 732株嗜血杆菌 ,采用Ｋ Ｂ纸片扩散法 ,对阿奇霉素做了药敏试验 ,并与其他 5种抗菌药物作了比较 ,报告如下。1　材料与方法1 1　菌种嗜血杆菌 732株 ,其中流感嗜血杆菌 30 2株 ,占41 3% ;副流感嗜血杆菌 430株 ,占 5 8 7%。1 2　培养基　巧克力培养基按《全国临床检验操作规程》(1997年第 2版 )由本室自制 ;血琼脂和Ｍ Ｈ琼脂 (Ｏｘｏｉｄ)作鉴定用 ;ＨＴＭ培养基 (Ｏｘｏｉｄ)作药敏用。1 3　药敏纸片　阿奇霉素、头孢噻肟、头孢呋辛、…     

嗜血杆菌分离培养基的评价与应用

目的：选择合适的嗜血杆菌分离培养提高嗜血杆菌分离率。方法：将嗜血杆菌接种于7种培养基上,计算和比较7种培养基上嗜血杆菌的平均生长指数（GI）;对325份呼吸道标本进行检测,比较巧克力琼脂（BRchoc）和加万古霉素巧克力琼脂(BRchoc-V）的嗜血杆菌分离率。结果：添加2％鲜酵母浸液和50％鲜牛肉浸液的各种培养基,较未加者的GI值明显增高;兔血巧克力琼脂培养基较羊血巧克力琼脂培养基的GI值为高（P＜0.001）;加万古霉素（50μg/ml）的兔血巧克力琼脂培养基,较未加者对嗜血杆菌的分离率（77.2％／32.0％）明显提高（P＜0.001）。结论：培养基中增加鲜酵母浸液等营养物质可促进流感嗜血杆菌的生长,制培养基所用血源以兔血最佳,其次为羊血;分离培养基加入50μg/ml的万古霉素,可显著提高嗜血杆菌的分离率。     

嗜血杆菌2种分离培养基的比较

嗜血杆菌是一种只侵犯人类的革兰阴性杆菌,存在于上呼吸道中,是引起急性呼吸道传染病的主要病原菌之一,也可侵入血液继发多种感染性疾病,如脑膜炎、肺炎、骨髓炎等。嗜血杆菌的分离对临床上早期诊断和治疗极为重要,由于该菌生长时需要X或/和V因子,不易检出,因此选择一种配制方法简单且适合该菌生长的培养基来提高分离率显得尤为关键,我们使用万古巧克力培养基,明显提高了嗜血杆菌的分离率。一、材料和方法1 .培养基　巧克力培养基:用胰酶—大豆琼脂(法国生物梅里埃公司产品)作基础,按用量加入蒸馏水,1 1 5°C1 5min高压灭菌后,冷却至80°C…     

Rapid determination of X/V growth requirements of Haemophilus species in broth

A broth system was developed for rapid identification of the requirement for X factor (hemin), or V factor (NAD), or both for growth of Haemophilus species. This system was compared to growth around paper discs/strips impregnated with factors X and/or V. The broth system consisted of three tubes, each containing brain-heart infusion broth supplemented with V factor, X factor, or both. Each tube was inoculated with a saline suspension of an Haemophilus isolate, and the broths were shaken for aeration at 37 degrees C. Under these conditions turbidity or clumping was usually evident after 4-5 hr only in the broth(s) containing the required supplement(s). A few strains requiring only V factor required overnight incubation. One hundred fifty-six Haemophilus isolates were tested for growth around supplemented discs/strips or in supplemented broths: 129 were H. influenzae/aegypticus, 25 were of various species that required only V factor, and 2 were H. aphrophilus. Ten of 89 H. influenzae isolates from the respiratory tract were misidentified by satellitism. All isolates were correctly identified by growth in supplemented broths. The cost of the broth assay was about 60 cents/test, whereas the satellite assay cost about 120 cents/test. Serotyping and antibiotic sensitivity testing could be performed directly from the broth culture. Determination of X and/or V requirement by Haemophilus species with supplemented broths was sensitive, rapid, and inexpensive.     

尿路感染生物被膜菌临床分布及耐药性研究

目的筛选尿路感染生物被膜菌,了解其临床分布,进而通过生物被膜菌与浮游菌耐药差异性分析,探讨细菌生物被膜对抗生素的耐药特性,以及初探生物被膜菌在体内较“真实”的耐药性。方法收集120例导尿管相关尿路感染（UTIs）患者导尿管和尿液标本,筛选生物被膜菌株和相应浮游株,对生物被膜半定量检测,药敏实验分析生物被膜菌株和相应浮游菌在普通MH培养基上耐药性差异以及生物被膜阳性菌在泊洛沙姆（Poloxamer,F-127）培养基和普通MH培养基上耐药差异性的分析。结果120例UTIs患者导尿管中筛选出生物被膜菌48株,阳性率为40％;生物被膜菌与相应的浮游菌在普通MH培养基上药敏结果相似,差异无统计学意义（P〉0．05）;生物被膜菌在Poloxamer培养基和MH培养基上药敏结果差异有统计学意义（P〈0．05）,且前者耐药性更强。结论本院尿路感染生物被膜菌在临床上以葡萄球菌属和肠球菌属为主;生物被膜菌与浮游菌在体外耐药性分析未见明显差异,推测Poloxamer培养基有可能表现生物被膜菌较真实的对药物的耐受情况,其耐受性更强。     

An evaluation of methods of determining the hemolytic activity of Klebsiella in solid nutrient media]

Hemolytic activities of Klebsiella-237 strains of various origins were measured in solid nutrient media by three methods making use of six different agar bases for the preparation of blood nutrient media. The authors analyze the agar bases, discuss the advantages and shortcomings of all the methods used with consideration for the purpose of the investigation. The findings evidence a higher hemolytic activity in Klebsiella isolated from patients as against the hemolytic activities of reference strains.     

Selective culture media for Haemophilus bacteria

A selectivity factor was specified to the previously developed nutrient medium meant for the cultivation of Haemophilus bacteria--CAE, which is based on the acid hydrolysate of casein, enzyme hydrolysate of animal blood (aminopeptide) and an extract of nutrient yeast. The above nutrient medium containing additionally growth factors V and X, glucose and bacitracin was shown to fit well for the primary sowing of Haemophilus bacteria and it can be used in diagnostic examinations.     

FURTHER OBSERVATIONS CONCERNING GROWTH REQUIREMENTS OF HEMOPHILIC BACILLI

1. 19 strains of non-hemolytic hemophilic bacilli were studied. 17 required the addition of V and X growth accessory factors to their media, 2 required the addition of only V. 2. Of 15 strains of hemolytic hemophilic bacilli examined, 10 were found to require the addition of only V to their media, 3 the addition of V and X in the form of yeast extract and hematin, and 2 the addition of accessory growth factors in the form of blood. 3. In media to which neither V nor X had been added true symbiosis was found to occur on growing B. hemoglobinophilus canis, requiring the addition of X, with B. parainfluenzæ, requiring the addition of V, or with hemolytic strains of bacilli, requiring the addition of only V.     

消毒剂杀灭淋球菌的试验方法研究

淋球菌对营养要求特殊,普通培养基上生长较慢。为科学评价化学消毒剂对淋球菌的杀灭效果,采用平板浸润接种法对不同培养基进行比较,以评价杀菌效果的差异。结果,淋球菌在普通营养琼脂和酵母浸膏琼脂上生长的菌落均小于2 mm;在酵母浸膏血琼脂和巧克力琼脂上生长的菌落为2~3 mm,计数回收率高出26%。而平板涂布法的回收菌数低于平板浸润法,其标准差大于平板浸润法。进行悬液定量杀菌试验时,采用酵母浸膏血平板浸润接种方法。观察临床分离的淋球菌和标准菌株对化学消毒剂的抗力,选择抗力较高的标准菌株作有机干扰物试验。用大白兔作为淋球菌感染部位和感染量的试验动物。结果表明,用酵母浸膏血琼脂替代巧克力琼脂培养淋球菌无统计学差异,淋球菌用平板浸润法接种比平板涂布法好。临床分离菌株对化学消毒剂的抗力略低于标准菌株。在30 g/L牛血清白蛋白的存在下,达到消毒要求的消毒剂浓度是PBS菌悬液的消毒剂浓度的5~6倍。淋球菌可通过皮肤、角膜感染大白兔的泌尿系统,可感染角膜菌量为80 cfu。故建议在30 g/L牛血清白蛋白的条件下,淋球菌经消毒剂处理后,下降5个对数值,作为消毒合格标准较可靠。     

进口培养基对卡他布拉汉菌分离鉴定的评价

目的了解呼吸道感染病人痰中卡他布拉汉菌的分离率方法采用法国生物梅里埃公司生产的哥伦比亚琼脂和DNA酶琼脂分离痰中的卡他布拉汉菌和做关键性的生化反应,鉴定结果与APINH试条进行对照。同时用国产培养基做对比试验结果卡他布拉汉菌在哥伦比亚琼脂配制的血平板上菌落明显大于国产血平板,与非致病性奈瑟氏菌有显著差别。关键性的生化反应在DNA酶琼脂平板上均显示阳性。鉴定结果的准确性与APINH试条完全相同。国产的DNA酶微量生化管仅4．4％为阳性。结论哥伦比亚琼脂和DNA酶琼脂是分离鉴定痰中卡他布拉汉菌良好的试剂。结合痰涂片镜检见到大量脓细胞吞噬G-双球菌的现象,我科在98年409例痰中共分离出45例卡他布拉汉菌,分离率为11％。占痰中各类病原菌的第4位。     

两种培养基的细菌动力检测结果准确性比较

目的：寻求一种对细菌动力试验结果更容易判定的半固体培养基。方法：在传统半固体培养基中加入氯化三苯四氮唑（TTC）进行动力试验,并与传统半固体培养基比较,分析这两种半固体培养基结果的相关性和准确性。结果：198株试验菌中,动力阳性株143株,动力阴性株55株。传统半固体培养基对动力阳性菌符合率为90．9％,对动力阴性菌符合率为100．0％;TTC半固体培养基对动力阳性菌符合率为98．6％,对动力阴性菌符合率为96．3%。统计分析表明,用TTC半固体培养基进行细菌动力实验与传统培养基有较好的相关性（x^2=141．0,P〈0．005）,且比传统半固体培养基结果更容易判断,准确性更高。结论：TTC半固体培养基可用于常规细菌动力鉴定。     

Effect of media composition on the susceptibility of Xanthomonas maltophilia to beta-lactam antibiotics.

The susceptibility of Xanthomonas maltophilia strains to beta-lactams was shown to depend on the concentrations at which individual media were prepared. MIC and disc susceptibility tests were performed on solidified Mueller-Hinton and Iso-Sensitest media prepared at 0.1, 0.3, 1, and 3 x the concentrations recommended by the manufacturers. Nine of ten X. maltophilia strains tested showed increasing sensitivity to meropenem, cefotaxime, cefoperazone, piperacillin and latamoxef as the nutrient concentrations of the two media were increased. The opposite pattern was found with one strain (NCTC 10257) on Mueller-Hinton agar. This strain behaved inconsistently on Iso-Sensitest agar. Previous studies have shown that medium-dependent susceptibility in X. maltophilia is not related to beta-lactamase expression. The present study demonstrated that 22 and 25 kDa outer membrane proteins were induced on media with higher nutrient concentrations. The possible relationship of these proteins to sensitivity is considered.     

Identification of beta-lactamase-negative, ampicillin-resistant strains of Haemophilus influenzae with four methods and eight media

A challenge set of 143 non-beta-lactamase-producing strains of Haemophilus influenzae was tested for ampicillin susceptibility on two broth media and six agar media, using broth microdilution, agar dilution, disk diffusion, and E-test procedures. When beta-lactamase-negative, ampicillin-resistant (BLNAR) strains were defined as those for which the ampicillin MIC was > or = 4.0 microg/ml, 5 to 44% of our selected strains were BLNAR depending on the medium and/or test method used. If nonsusceptible strains for which ampicillin MICs were intermediate were included in the BLNAR category, 32 to 50% of our isolates would be considered BLNAR. These data emphasize the need for a standardized testing procedure and a universal definition of BLNAR strains before the clinical relevance of such strains can be evaluated. NCCLS dilution tests with haemophilus test medium broth or agar are preferred for testing ampicillin against H. influenzae.