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改良的KANE/FISHER系统对常见皮肤癣菌的鉴定 总被引:1,自引:0,他引:1
目的 以改良的KANE/FISHER系统对常见皮肤癣菌进行鉴定,探索建立一个临床实用的标准化皮肤癣菌鉴定系统.方法 选取KANE/FISHER鉴定系统中的6种主要培养基对皮肤癣菌ATCC株和经初步鉴定的临床株共63株进行培养鉴定.结果 改良的KANE/FISHER鉴定系统基本可以将实验菌株鉴定到种的水平.结论 改良的KANE/FISHER鉴定系统对常见皮肤癣菌能做到种的鉴定,可作为标准化鉴定方法在实验室中应用. 相似文献
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目的报告国内首例多育赛多孢菌致鼻窦炎,并探讨致病菌的鉴定及其对抗真菌药物体外敏感性。方法取患者左侧上颌窦分泌物进行真菌培养和形态学鉴定,分离菌株β-球蛋白、rDNAITS序列分析确切鉴定,对分离菌进行7种抗真菌药体外药敏试验。结果根据菌株的形态学特点和基因序列结果鉴定为多育赛多孢菌。体外药敏试验显示该菌对7种抗真菌药物耐药。结论多育赛多孢菌所致的真菌病较少见,其确切鉴定靠形态学特征和基因分析。该菌株对多种抗真菌药物耐药。 相似文献
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目的 了解大连地区感染性腹泻病原菌的种类和流行特征,为感染性腹泻的预防控制和临床治疗提供依据.方法 采用常规粪便培养方法,对分离菌做血清学分型,用MicroScan WalkAway-40全自动细菌鉴定及药敏分析仪对分离菌做生化鉴定,对检出的志贺菌做药敏试验,并用WHONET 5.4软件对药敏结果进行统计分析.结果 监测872例腹泻患者样本,检出感染性腹泻病原菌109株,阳性率为12.5%,其中志贺菌所占比例最大,其次为沙门菌,副溶血弧菌排第三.志贺菌对碳青霉烯类、氟喹诺酮类和头孢菌素类等敏感,对青霉素类和复方磺胺类耐药.结论 志贺菌是大连地区感染性腹泻的首要病原菌,沙门菌和副溶血弧菌次之,应依据此监测结果做好大连地区腹泻病的防治工作;同时根据耐药性监测结果合理使用抗生素应对志贺菌,减少耐药菌株出现. 相似文献
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《中国真菌学杂志》2016,(1)
目的了解本院门诊拟诊皮肤真菌病患者真菌感染状况,掌握近期本地区皮肤真菌感染菌种分布结构。方法采用KOH直接涂片或染色显微镜观察与分离培养相结合的方法,对分离的菌落进行形态学鉴定,对真菌镜检和培养鉴定结果进行统计分析。结果 47 766例拟诊皮肤真菌病患者中,直接镜检和(或)培养阳性25 078例,占52.50%。其中皮肤癣菌病9 310例(48.94%)、马拉色菌感染8 938例(46.99%)、念珠菌及其他酵母菌感染603例(3.17%)。除外马拉色菌镜检和(或)培养阳性,其余38 839例镜检阳性率40.33%、分离培养阳性率25.99%。分离培养与镜检阳性符合率61.41%。皮肤癣菌菌种结构方面,红色毛癣菌为优势菌种占88.19%。结论皮肤癣菌病位居皮肤真菌病之首,以红色毛癣菌为优势菌,念珠菌及其他酵母菌感染亦值得关注。 相似文献
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目的了解食源性相关腹泻非伤寒沙门菌感染的菌种分布及其对抗菌药物敏感性,为控制该类细菌的感染及传播提供技术支持。方法对2015-2017年本系统2家社区卫生服务中心就诊的食源性相关腹泻患者粪便(或肛拭子)标本采用直接分离与增菌分离相结合的方法常规培养分离获得42株沙门菌;采用血清凝集法进行快速血清分型,并进行自动化生化鉴定及抗菌药物敏感性试验;通过现况调查进行流行病学危险因素分析。结果自动化生化鉴定结果能够覆盖常规血清学快速鉴定结果,同属沙门菌群;42株沙门菌以肠炎沙门菌、鼠伤寒沙门菌和斯坦利沙门菌为主,其中肠炎沙门菌占全部菌株的23.81%,鼠伤寒沙门菌占19.06%,斯坦利沙门菌占14.29%;其中肠炎沙门菌可对氟喹诺酮类、三四代头孢菌素类与碳青霉烯类等敏感(敏感率可达97.00%以上),而对氨基糖苷类可产生双向耐药。通过现况调查,发现患者有腹痛、腹泻等胃肠道症状,可伴有发热,所有患者48h内有可疑食物暴露史,无水源性案例,均为散发。结论菌种鉴定应以常规快速血清学凝集结果为准,自动化生化鉴定仅供参考,并将鉴定菌种与药敏报告相关联,可根据药敏结果合理选用敏感抗菌药物;应对社区居民开展针对性的健康宣教。 相似文献
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Sue T Obolonkin V Griffiths H Villas-Bôas SG 《Applied and environmental microbiology》2011,77(21):7605-7610
The early detection of microbial contamination is crucial to avoid process failure and costly delays in fermentation industries. However, traditional detection methods such as plate counting and microscopy are labor-intensive, insensitive, and time-consuming. Modern techniques that can detect microbial contamination rapidly and cost-effectively are therefore sought. In the present study, we propose gas chromatography-mass spectrometry (GC-MS)-based metabolic footprint analysis as a rapid and reliable method for the detection of microbial contamination in fermentation processes. Our metabolic footprint analysis detected statistically significant differences in metabolite profiles of axenic and contaminated batch cultures of microalgae as early as 3 h after contamination was introduced, while classical detection methods could detect contamination only after 24 h. The data were analyzed by discriminant function analysis and were validated by leave-one-out cross-validation. We obtained a 97% success rate in correctly classifying samples coming from contaminated or axenic cultures. Therefore, metabolic footprint analysis combined with discriminant function analysis presents a rapid and cost-effective approach to monitor microbial contamination in industrial fermentation processes. 相似文献
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Vahid Molla Kazemiha Amir Amanzadeh Arash Memarnejadian Shahram Azari Mohammad Ali Shokrgozar Reza Mahdian Shahin Bonakdar 《Cytotechnology》2014,66(5):861-873
Mycoplasma contamination in cell culture is considered as serious problem in the manufacturing of biological products. Our goal in this research is to find the best standard and rapid method with high sensitivity, specificity, accuracy and predictive values of positive and negative results for detection of mycoplasma contamination in cell cultures of the National Cell Bank of Iran. In this study, 40 cell lines suspected to mycoplasma contamination were evaluated by three different methods: microbial culture, enzymatic mycoalert® and molecular. Enzymatic evaluation was performed using the mycoalert® kit while in the molecular technique, a universal primer pair was designed based on the common and fixed 16SrRNA ribosomal sequences used. Mycoplasma contaminations in cell cultures with molecular, enzymatic and microbial culture methods were determined as 57.5, 52.5 and 40 %, respectively. These results confirmed the higher rate of sensitivity, specificity and accuracy for the molecular method in comparison with enzymatic and microbial methods. Polymerase chain reaction (PCR) assay based on fixed and common sequences in the 16SrRNA, is a useful valuable and reliable technique with high sensitivity, specificity and accuracy for detection of mycoplasma contamination in cell cultures and other biological products. The enzymatic mycoalert® method can be considered as a substitution for conventional microbial culture and DNA staining fluorochrome methods due to its higher sensitivity, specificity and speed of detection (<20 min). 相似文献
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Baumstummler A Chollet R Meder H Rofel C Venchiarutti A Ribault S 《Letters in applied microbiology》2010,51(6):671-677
Aims: Microbial contamination of cell culture production processes is a current concern for biopharmaceutical industries. Traditional testing methods require several days to detect contamination and may advantageously be replaced by a rapid detection method. We developed a new method combining membrane filtration to microcolonies fluorescence staining method (MFSM) and compared it to epifluorescence microscopy. Methods and Results: Both methods were used to detect bacteria in CHO cells cultures. The epifluorescence microscopy showed to be limited by filterability, media interference and nonrobustness issues, whereas MFSM enabled consistent detection of Bacillus cereus, Staphylococcus epidermidis and Propionibacterium acnes after, respectively, 8, 9 and 48 h of incubation. Thanks to the nondestructive feature of the MFSM, stained membranes could be reincubated on culture media to yield visible colonies used for identification. Conclusions: The new method described in this study showed its ability to detect microbial contaminants in cell culture samples with time‐to‐results from 2–5 times shorter than the traditional testing method. Significance and Impact of the Study: The MFSM can be used as monitoring tool for cell cultures to significantly shorten detection times of microbial contamination, while preserving the ability to identify the contaminants and their viability. 相似文献
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Mycoplasma is well recognized as one of the most prevalent and serious microbial contaminants encountered within the manufacturing of biopharmaceuticals from the research phase to clinical development and production. The potential for mycoplasma contamination within cell culture systems was first identified by Robinson et al. in 1956 [1]. Presently, contamination rates in established cell cultures have been reported between 15 and 35% with considerably higher occurrence cited in certain selected populations [2]. In the last few years, there has been an expansion of diagnostic approaches for mycoplasma detection with the development and validation of rapid microbiological methods. The objective of this study was to determine current levels of mycoplasma infection of cell cultures, cell substrates and biologicals within a client based population. Retrospective comparison of 40,000 sample results was done to determine total contaminations rates amongst four (4) individual analytical assays. The establishment of reference data, such as existing contamination rates, becomes important in the critical appraisal of rapid microbiological methods for the detection of mycoplasma. 相似文献
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Vahid Molla Kazemiha Shahin Bonakdar Amir Amanzadeh Shahram Azari Arash Memarnejadian Shirin Shahbazi Mohammad Ali Shokrgozar Reza Mahdian 《Cytotechnology》2016,68(4):1063-1080
Mycoplasmas are the most important contaminants of cell cultures throughout the world. They are considered as a major problem in biological studies and biopharmaceutical economic issues. In this study, our aim was to find the best standard technique as a rapid method with high sensitivity, specificity and accuracy for the detection of mycoplasma contamination in the cell lines of the National Cell Bank of Iran. Thirty cell lines suspected to mycoplasma contamination were evaluated by five different techniques including microbial culture, indirect DNA DAPI staining, enzymatic mycoalert® assay, conventional PCR and real-time PCR. Five mycoplasma-contaminated cell lines were assigned as positive controls and five mycoplasma-free cell lines as negative controls. The enzymatic method was performed using the mycoalert® mycoplasma detection kit. Real-time PCR technique was conducted by PromoKine diagnostic kits. In the conventional PCR method, mycoplasma genus-specific primers were designed to analyze the sequences based on a fixed and common region on 16S ribosomal RNA with PCR product size of 425 bp. Mycoplasma contamination was observed in 60, 56.66, 53.33, 46.66 and 33.33 % of 30 different cell cultures by real-time PCR, PCR, enzymatic mycoalert®, indirect DNA DAPI staining and microbial culture methods, respectively. The analysis of the results of the different methods showed that the real-time PCR assay was superior the other methods with the sensitivity, specificity, accuracy, predictive value of positive and negative results of 100 %. These values were 94.44, 100, 96.77, 100 and 92.85 % for the conventional PCR method, respectively. Therefore, this study showed that real-time PCR and PCR assays based on the common sequences in the 16S ribosomal RNA are reliable methods with high sensitivity, specificity and accuracy for detection of mycoplasma contamination in cell cultures and other biological products. 相似文献
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Mycoplasma contamination of cell cultures: Incidence, sources, effects, detection, elimination, prevention 总被引:11,自引:0,他引:11
The contamination of cell cultures by mycoplasmas remains a major problem in cell culture. Mycoplasmas can produce a virtually
unlimited variety of effects in the cultures they infect. These organisms are resistant to most antibiotics commonly employed
in cell cultures. Here we provide a concise overview of the current knowledge on: (1) the incidence and sources of mycoplasma
contamination in cell cultures, the mycoplasma species most commonly detected in cell cultures, and the effects of mycoplasmas
on the function and activities of infected cell cultures; (2) the various techniques available for the detection of mycoplasmas
with particular emphasis on the most reliable detection methods; (3) the various methods available for the elimination of
mycoplasmas highlighting antibiotic treatment; and (4) the recommended procedures and working protocols for the detection,
elimination and prevention of mycoplasma contamination. The availability of accurate, sensitive and reliable detection methods
and the application of robust and successful elimination methods provide powerful means for overcoming the problem of mycoplasma
contamination in cell cultures.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
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Meruva NK Penn JM Farthing DE 《Journal of industrial microbiology & biotechnology》2004,31(10):482-488
Several microbial volatile organic compounds (MVOCs) that can serve as potential chemical markers for microbial contamination in tobacco have been identified. Four different fungal species, Aspergillus niger (AN), A. ornatus (AO), Pencillium chrysogenum (PC) and Rhizopus stolonifer (RS), commonly reported in moldy tobacco were cultured and screened for MVOCs. Because the MVOCs emitted by a microbial species are substrate specific, the fungal strains were separately grown on potato dextrose agar (PDA) and tobacco products. MVOCs from the mold cultures grown on PDA and tobacco products were extracted using closed-loop stripping analysis (CLSA) and identified by gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS). Some of the prominent tobacco mold markers identified by this method include: 1-octen-3-ol; 2-octen-1-ol; 2-methyl-1-butanol; 3-methyl-1-butanol; 1-octene and 2-pentanone. In particular, 1-octen-3-ol was detected in all the mold cultures and moldy tobacco samples analyzed. Olfactory evaluation of 1-octen-3-ol indicated a characteristic musty odor and the odor threshold was determined to be approximately 200 ng/ml. The limits of detection for 1-octen-3-ol using GC/TOF-MS and GC/mass selective detector (MSD) in the full-scan mode and selected ion monitoring (SIM) mode were investigated. The CLSA-GC/TOF-MS demonstrates a fast, sensitive and semi-quantitative analytical technique for screening tobacco materials for the presence of mold via chemical markers of microbial contamination. 相似文献
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Mycoplasma contamination in cell culture is a serious setback for the cell-culturist. The experiments undertaken using contaminated cell cultures are known to yield unreliable or false results due to various morphological, biochemical and genetic effects. Earlier surveys revealed incidences of mycoplasma contamination in cell cultures to range from 15 to 80%. Out of a vast array of methods for detecting mycoplasma in cell culture, the cytological methods directly demonstrate the contaminating organism present in association with the cultured cells. In this investigation, we report the adoption of a cytological immunofluorescence assay (IFA), in an attempt to obtain a semi-automated relative quantification of contamination by employing the user-friendly Photoshop-based image analysis. The study performed on 77 cell cultures randomly collected from various laboratories revealed mycoplasma contamination in 18 cell cultures simultaneously by IFA and Hoechst DNA fluorochrome staining methods. It was observed that the Photoshop-based image analysis on IFA stained slides was very valuable as a sensitive tool in providing quantitative assessment on the extent of contamination both per se and in comparison to cellularity of cell cultures. The technique could be useful in estimating the efficacy of anti-mycoplasma agents during decontaminating measures. 相似文献
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Keeping sterile stocks or cultures of microalgae is fundamental to microalgae biotechnology as well as basic scientific research. However, contamination by bacteria and/or fungi in microalgae cultures or stocks is often a problem. Here, we have developed a strategy for reducing or eliminating bacterial and fungal contamination by using a cocktail of antibiotics. Chlamydomonas reinhardtii P. A. Dang., a widely used unicellular green alga, has been used as a testing organism. A combination of ampicillin, cefotaxime, and carbendazim removed or reduced contamination by three different bacteria and two different fungi tested. A step‐by‐step procedure is provided, which is simple, economical, and effective. 相似文献
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Kiyoshi Tsuji E. M. Stapert John H. Robertson Peter M. Waiyaki 《Applied microbiology》1970,20(5):798-801
A sensitive sterility testing procedure for the detection of microbial contamination in petrolatum-based ointments is described. The method involves dissolving the ointment in filter-sterilized isopropyl myristate and filtering through a membrane filter. Improved sensitivity is obtained by blending the membrane in Trypticase Soy Broth before incubation. Filter-sterilized isopropyl myristate is shown to be less toxic to microorganisms than heat-sterilized isopropyl myristate. The isopropyl myristate method is more sensitive than the polyethylene glycol-ether method for the detection of microbial contamination. 相似文献