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目的 应用多种方法对CMCC(B) 32220鉴定和分析,了解其全基因组的分子特征。方法 将待鉴定菌株CMCC(B) 32220复苏培养后,依次进行形态学、全自动生化鉴定、基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析、16S rRNA基因以及全基因序列测定,应用生物信息学方法分析其基因组基本特征,并预测基因功能、耐药性及致病性。结果 经全自动生化和MALDI-TOF-MS鉴定分析菌株CMCC(B) 32220均为铅黄肠球菌。CMCC(B) 32220菌株与NCBI数据库中已发表2株代表性铅黄肠球菌16S rRNA基因序列相似度分别为99.4%和98.9%,而与不同肠球菌16S rRNA基因序列相似度为94.8%~98.7%。CMCC(B) 32220染色体全基因组序列全长为3 195 952 bp,包含3 054个基因,其中具有直系同源簇(COG)功能2 453个,参与KEGG代谢通路1 622个。而基于全基因组水平的平均核苷酸一致性(ANI)分析证实CMCC(B) 32220菌株与已发表的铅黄肠球菌的ANI值为95.1%,而与其他3株肠球菌属不同种(屎肠球菌、鸡肠... 相似文献
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肠球菌属分类和鉴定 总被引:1,自引:0,他引:1
马俊春 《上海医学检验杂志》1991,6(4):249-252
一、肠球菌属的分类体系肠球菌分类系统近年来有较大变化,以往将肠球菌归入链球菌属,并把D群链球菌分成肠球菌和非肠球菌。随着现代细菌分类技术发展,1984年把原来的链球菌属分为3个菌属:即链球菌属(Streptococcus),肠球菌属(Enterococcus)和乳球菌属 相似文献
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肠球菌分类与鉴定新进展 总被引:21,自引:0,他引:21
肠球菌属(Enterococcus)细菌为兼性厌氧、触酶阴性、链状排列的革兰染色阳性球菌,其DNA的G C含量为(37- 45)mol%。多数菌种产生D抗原,有些菌种可产生Q抗原。为人和动物胃肠道正常菌群之一,可引起抵抗力低下宿主的多种机会感染,现已成为医院感染的重要致病菌。1分类肠球菌最初被分类为肠内的革兰阳性球菌,之后归入链球菌属。1930年,随着Lancefield血清分型系统的建立,肠球菌被分类为D群链球菌,并可应用不同的生理生化特性区别于非肠球菌D群链球菌,如牛链球菌。由于该群菌10℃、45℃、65g/L NaCl中均生长,耐受60℃30 min,并在40%胆汁 相似文献
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李金钟 《中华检验医学杂志》2004,27(7):465-467
1989年有关文献报道,耳炎差异球菌被从慢性中耳炎患者的中耳渗出液(media ear effusion,MEE)中分离,并对这些细菌的生物学特性进行了描述,认为这些细菌是在低G C含量革兰阳性球菌中,但 相似文献
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204株肠球菌鉴定及其药敏试验研究 总被引:9,自引:0,他引:9
由于肠球菌对广谱抗菌药物耐药 ,对高浓度氨基糖甙类抗生素及万古霉素的耐药性的流行 ,医院感染率呈逐年上升趋势。笔者根据马俊春等[1] 的鉴定肠球菌的方法 ,并对其耐药性进行分析。菌株 :从临床分离肠球菌 2 0 4株 ,其中从尿液标本分离10 0株 ,咽拭子 2 7株 ,手术伤口分泌物 2 5株 ,腹腔积液 14株 ,胆汁 13株 ,非手术伤口分泌物 9株 ,宫颈、阴道分泌物 7株 ,胸腔积液 4株 ,血液 3株 ,脑脊液 2株。标准菌株为ATCC2 5 92 3(金黄色葡萄球菌 )、ATCC2 92 12 (粪肠球菌 )购自卫生部临床检验中心。药敏试验 :采用M H琼脂平板培养基 ,… 相似文献
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肠球菌的分离鉴定及抗生素敏感性试验 总被引:1,自引:0,他引:1
肠球菌的分离鉴定及抗生素敏感性试验孙延波,张学英,黄红兰,关显智(白求恩医科大学微生物学教研室,长春130021)李淑华(长春医学高等专科学校微生物学教研室)关键词肠球菌属,抗生素,药物敏感试验肠球菌属细菌在自然界分布广,是人及动物肠道中的正常菌群,... 相似文献
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2002年,Shankar等在临床致病粪肠球菌中鉴定出第一个肠球菌致病岛,该岛大约150kh大小,有129个开放阅读框(ORFs),携带致病所需基因和潜在毒力基因。粪肠球菌PAI的可塑性非常强,其基因丢失的频率和可能机制目前尚不清楚。2004年Leavis等在屎肠球菌中也发现了PAI类似结构。 相似文献
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本文报告100株肠球菌系统生理生化,及药敏试验研究。其结果为:粪肠球菌89株,屎肠球菌11株。对其进行青霉素,氨苄青霉素,左氧氟沙星和呋喃妥因药敏试验,敏感率较高,并发现屎场球菌比粪肠球菌更耐药。对严重的肠球菌感染,在测定肠球菌高浓度氨基糖苷类耐药性指导下,以联合用药为佳. 相似文献
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克罗诺杆菌,原名阪崎肠杆菌,是污染婴幼儿配方奶粉的主要致病菌,能感染新生儿,并导致严重的坏死性小肠结肠炎、菌血症和脑膜炎等疾病。随着对该菌认识的深入,阪崎肠杆菌被命名为肠杆菌科独立的新属——克罗诺杆菌属。最新分类表明克罗诺杆菌属共包含7个种,每个种之间基因组构成和致病性存在差异,因此分种是克罗诺杆菌研究的基础,对进一步认识和了解克罗诺杆菌具有重要意义。近年来,除了传统生化方法分种,一系列分子生物学方法也应用于克罗诺杆菌的分种。本文综述了克罗诺杆菌的生物学特性和目前用于克罗诺杆菌分种的各种方法,以期增加对克罗诺杆菌分种方法的认识,为今后的研究工作提供依据。 相似文献
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We have developed PCR and Multiplex PCR assays for the detection of medically important Candida spp. using different species and genus-specific PCR primers selected within the MP65 gene, a recently cloned gene encoding a mannoprotein adhesin. The genus-specific PCR primers were able to amplify Candida species DNA (100% positivity) whereas DNA from all other isolates tested, belonging to other fungal genera, was not amplified. The species-specific PCR primers allowed differentiation of each of five Candida species by the amplicon length produced. No amplicons were detected using species- or genus-specific primers in several bacterial or human DNA templates. The methods described in this study are reproducible, simple and specific. The total time required for each PCR method was less than 4 h from the extraction to the visualized amplicons after PCR. In conclusion, we developed PCR methods to differentiate the five most medically important Candida species using primers directed to the MP65 gene. 相似文献
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Development of a Salmonella-specific biotinylated DNA probe for rapid routine identification of Salmonella 总被引:6,自引:0,他引:6
J. M. Gopo R. Melis E. Filipska R. Meneveri J. Filipski 《Molecular and cellular probes》1988,2(4):271-279
Classical microbiological techniques used in the detection and identification of Salmonella spp. in foods, drinking water and clinical samples are relatively lengthy. Immunoassays, on the other hand, have the major disadvantage of often generating false positives and false negatives. Recombinant DNA technology offers more efficient alternatives to the detection of a specific organism by employing cloned DNA sequences unique to the organism. Demonstration of a presence of complementary sequences among a heterogeneous population of molecules of DNA isolated from bacteria can be made by using a DNA-DNA hybridization technique. We have obtained a fragment of DNA from Salmonella typhimurium chromosomal DNA, cloned it in Escherichia coli plasmid and tested it in colony hybridization tests with 57 strains of Salmonella and other enterobacteriaceae. In all tests, the fragment was found to be Salmonella-specific in that it gave a positive reaction with all strains of Salmonella tested and was negative when tested against other Enterobacteriaceae. 相似文献
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Po-Chi Soo Yu-Tze Horng Kai-Chih Chang Jann-Yuan Wang Po-Ren Hsueh Chun-Yu Chuang Chia-Chen Lu Hsin-Chih Lai 《Molecular and cellular probes》2009,23(5):240-246
We had previously developed a nested polymerase chain reaction (PCR)–immunochromatography test (ICT) for identification of Mycobacterium tuberculosis (MTB) and differentiation of MTB from other members of M. tuberculosis complex (MTBC) from clinical sputum samples (Soo P.C. et al., Journal of Microbiological Methods. 2006, 66(3):440–8.). To further improve the detection flexibility, simplicity and efficiency, and reduce the cost, in this study, an alternative molecular diagnosis assay that utilizes gold nanoparticles derivatized with thiol modified oligonucleotides was developed. The gold nanoparticles probes, GP-1/GP-2 for IS6110 and GP-3/GP-4 for Rv3618, were designed to specifically hybridize with target DNAs of MTBC and MTB strains, respectively. Efficacy of the gold nanoparticle probes assay was evaluated by directly and simultaneously detecting not only MTBC but also MTB from 600 clinical sputum specimens. Results were compared with traditional culture and biochemical identification methods together with patients' clinical assessments. This assay showed a 96.6% sensitivity and 98.9% specificity towards detection of MTBC, and a 94.7% sensitivity and 99.6% specificity for detection of MTB. In conclusion, the gold nanoparticle probes assay is a simple, rapid, cost-effective and accurate detection system and shows great potential in clinical application of MTBC and MTB detection, especially in developing countries. 相似文献
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目的 建立一种检测胎儿弯曲菌胎儿亚种、性病亚种和龟亚种的多重PCR检测方法。方法 使用针对胎儿弯曲菌、性病亚种和龟亚种的特异性引物,优化反应体系及反应条件,使用38株菌株(18株胎儿弯曲菌和20株非胎儿弯曲菌)验证其特异性,并将该方法应用于53份爬行动物粪便筛查。结果 针对3种亚种均可扩增相应的片段,胎儿亚种只有1条359 bp大小的条带、性病亚种有2条分别为359 bp和156 bp大小的条带,龟亚种有2条分别为359 bp和266 bp大小的条带。优化后的最佳扩增条件:退火温度58℃,引物MG3f和Cf359r浓度为0.1μmol/L,ISC2mf和ISC2mr、CoA266f和CoA266r的浓度0.2μmol/L。对胎儿亚种、性病亚种和龟亚种的检出限分别为0.40 ng/μL、0.39 ng/μL和0.47 ng/μL。使用18株胎儿弯曲菌和20株非胎儿弯曲菌其特异性为100%。对53份爬行动物粪便进行筛查中有1份龟亚种阳性并分离出了相应的菌株。结论 本研究建立的多重PCR方法能利用1个反应体系同时鉴别3种亚种,从而为菌株快速鉴定、流行病学调查和溯源研究提供技术支持。 相似文献
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Henchal EA Teska JD Ludwig GV Shoemaker DR Ezzell JW 《Clinics in Laboratory Medicine》2001,21(3):661-678
The authors present an integrated approach for the identification of biological threat agents. The methods used have been used extensively in field exercises and during response to incidents of biological terrorism. A diagnostic system, which integrates the clinical diagnosis or medical intelligence with immunodiagnostic tests, rapid gene amplification assays, and standard culture, provides results of the highest quality and confidence. In the future, selected reagents and technologies will be distributed through a network of civilian and military laboratories. 相似文献