Infection with Nocardia species in Queensland. A review of 102 clinical isolates.

OBJECTIVE: To review the species distribution, pathologic significance and disease associations of clinical isolates of Nocardia and related bacteria in Queensland, and to examine the characteristics, treatment and outcome of patients infected with these organisms. DESIGN AND SETTING: A retrospective review of Queensland State Health Laboratory records provided microbiological data for Nocardia isolates referred from other laboratories during the period January 1983 to December 1988. Clinical information was extracted from hospital case notes, or obtained from detailed questionnaires completed by attending physicians. Nocardia isolates were classified as "significant" if specific treatment for nocardiosis was given, or on the basis of autopsy findings. PATIENTS: One hundred and two patients had a Nocardia species or a related organism isolated from clinical specimens during the study period. RESULTS: The 102 isolates included Nocardia asteroides (45), N, brasiliensis (35), N. caviae (5) and N. transvalensis (5). Clinical results were available for 93 patients, of whom 74 (80%) had a significant isolate recovered. Primary pulmonary or disseminated disease occurred in 35 patients, and was caused mainly by N. asteroides. Significant infections of skin and soft tissues, primarily due to N. brasiliensis, were found in 39 patients. Preexisting lung disease and treatment with steroids and immunosuppression were risk factors for pulmonary and disseminated nocardiosis. A history of inoculation in an outdoor setting was frequent in patients with cutaneous disease. Antibiotic regimens that included trimethoprim-sulfamethoxazole or another sulfonamide agent were used to treat the majority of patients with significant infection. Deaths were confined to those with pulmonary and disseminated disease, with a case fatality rate of 40% in that group. CONCLUSION: Infection with Nocardia species appears to be more common than is generally appreciated. The local species distribution and disease spectrum are similar to those described elsewhere. A high index of suspicion for nocardiosis should be maintained in susceptible hosts with pulmonary infiltrates, particularly when there is evidence for metastatic infection, and in patients with superficial infections and a history of outdoor injury.     

北京地区实验动物中嗜肺巴斯德杆菌的表型分析

目的对北京地区实验动物中的嗜肺巴斯德杆菌分离株进行表型分析,提高检测的准确性。方法通过生化鉴定和16S rDNA测序,对306株嗜肺巴斯德杆菌可疑菌株进行鉴定。结合分离株在血琼脂平皿上的菌落特征和生化特性,组成53个特征数据谱,进行表型特征的聚类分析。结果 306株分离株中,BD自动细菌鉴定系统和16S rDNA测序鉴定出嗜肺巴斯德杆菌的阳性率分别为164/306和227/306。227株嗜肺巴斯德杆菌真阳性的表型特征共有140种,Heyl型106种,Jawetz型23种。结论北京地区实验动物中嗜肺巴斯德杆菌主要为Heyl型感染,Jawetz型较少,表型特征多样,分布广泛。     

Nocardia asteroides osteomyelitis

Nocardia asteroides osteomyelitis has previously been described only in acutely ill patients. It may occur as an isolated event or part of the disease spectrum of disseminated nocardiosis. An 84 year old immune competent man with N. asteroides right tibial osteomyelitis had an unusual presentation, presumed mechanism of disease and outcome. His course affords an opportunity to review the literature on N. asteroides osteomyelitis and to discuss the unique features of his case.     

流感嗜血杆菌的蛋白电泳图谱分析及分型

目的 分析流感嗜血杆菌蛋白的成分及免疫原性。为流感嗜血杆菌菌苗的研制提供依据。方法 应用聚丙烯酰胺凝胶电泳分析５６株流感嗜血杆菌蛋白成分。结果 ５６株流感嗜血杆菌中不同分型流感嗜血杆菌４４株,占７９％。生物分型中,Ⅱ型为最高,占４８％,流感嗜血杆菌约有２０多条蛋白带,１１条为所有型别菌株共同具有的,其中以相对分子量为９６０００、５１０００及３１０００者含量较高,各菌株间的差异蛋白的相对分子量主要为     

PCR为基础的反向线点杂交及16S～23S rDNA间区测序分析5种少见类型奴卡菌

目的探讨5种新出现的奴卡菌种16S～23S rDNA内转录间隔区(internal transcribed spacer,ITS)DNA基因序列的多态性,筛查特异的奴卡菌种逆向线点杂交(reverse line blot,RLB)探针。方法对6例奴卡菌标本进行ITS基因扩增、克隆、测序,设计奴卡菌种特异探针及ITS间区rDNA基因特异的探针,进行以PCR为基础的RLB(PCR/RLB)杂交试验,不同的RLB型进行ITS基因测序分析,以鉴别不同的奴卡菌种及种内分型。结果发现2个Nocardia beijingensis菌株ITS间区有8个基因序列型,4个RLB型;N.thailandica有4个基因序列型及RLB型;N.blacklockiae有5个基因序列型,3个RLB型,其中N-bla RLBⅢ型与所有Nbla-ITS探针杂交没有信号;N.elegans有5个基因序列型,3个RLB型;N.vinacea有5个基因序列型,2个RLB型。结论通过PCR/RLB试验,准确的ITS基因测序认识了新出现的菌种,即N.beijingensis,N.blacklockiae,N.elegans,N.thailandica及N.vinacea。同时建立了可以大量筛查奴卡菌种的PCR/RLB方法,以进行快速临床诊断及流行病学研究。     

江苏汉族人群上颌前牙牙周生物型调查

目的：调查江苏汉族人群上颌前牙牙周生物型的分布,以及牙冠宽长比（crownwidth/crown length,CW/CL）、龈乳头高度（palilla height,PH）和附着龈宽度（attached gingival width,AGW）,为前牙美学区的口腔治疗提供解剖学参考。方法：招募江苏地区牙周健康的汉族志愿者108例（男52例,女56例）,根据牙周透射探针法,将上中切牙牙龈分成厚、薄两型,再结合牙齿形态和龈线起伏,细分为薄扇型、厚扇型或厚平型3种牙周生物型。同时,测量CW/CL、PH和AGW,分析上述解剖学指标在不同牙周生物型、性别、年龄和牙位间的分布差异。结果：江苏汉族人群中,薄扇型占30%,厚扇型占19%,厚平型占51%,平均CW/CL 0.84±0.09,平均PH（3.93±0.80）mm,平均AGW（4.80±1.39）mm。厚平型CW/CL和AGW明显大于其余两型（P＜0.05）,PH明显小于其余两型（P＜0.05）。结论：江苏汉族人群牙周生物型以厚平型为主,口腔治疗的美学风险相对较小,不同牙周生物型的CW/CL、PH和AGW存在差异。     

Primary cutaneous nocardiosis.

Seven patients with primary cutaneous nocardiosis were seen during a 20-month period. Six patients had a history of local trauma to an extremity or did frequent yard work. The spectrum of clinical disease included cellulitis, pustules,pyoderma, or lymphocutaneous syndrome mimicking sporotrichosis. Evidence of Nocardia infection elsewhere was absent, and no patient had a serious underlying disease. Nocardia brasiliensis was isolated from four cases, and N asteroides was identified in the other three patients. In two patients, disease resolved without chemotherapy, while a short course of a sulfonamide with or without trimethoprim therapy was successful in the remaining cases. The relatively benign nature of this disease, the usual failure to obtain cultures of superficial skin infections, and our accumulation of seven cases in less than two years suggest this disease is more common than previously recognized.     

马拉色菌对三种唑类抗真菌药的体外药敏试验

目的:探讨马拉色菌标准株和临床分离株对酮康唑、伊曲康唑和氟康唑的体外敏感性.方法:用生理生化及形态学方法对临床分离的马拉色菌菌株进行菌种鉴定.参照美国临床实验室标准委员会(NCCLS)制订的M27-A方案中酵母菌微量稀释法,检测7种马拉色菌标准株和42株临床分离株对酮康唑、伊曲康唑和氟康唑的最小抑菌浓度(MIC).结果:42株马拉色菌临床分离株包括5个种,分别是合轴马拉色菌、糠秕马拉色菌、钝形马拉色菌、球形马拉色菌、限制性马拉色菌.用M27-A方案中酵母菌微量稀释法测定3种唑类抗真菌药对7种马拉色菌标准株和42株临床分离株的敏感性由高到低分别为:伊曲康唑、酮康唑、氟康唑.结论:马拉色菌属的各菌种对酮康唑、伊曲康唑和氟康唑的敏感性存在差异;M27-A方案中酵母菌微量稀释法可用于马拉色菌的体外药敏试验.     

Diversity and frequency of Nocardia spp. in the soil of Isfahan province,Iran

Objective

To isolate and identify Nocardia spp. from soil in different regions of Isfahan province in the center of Iran.

Methods

This study was conducted in 32 districts (16 cities and 16 villages) in Isfahan province during two years. A total of 800 soil samples from these regions were studied by using kanamycin. The isolated Nocardia species were examined by gram and acid-fast staining and were identified biochemically and morphologically. The frequency and distribution of Nocardia spp. were determined in relation to different factors such as soil pH and temperate climate.

Results

From 153 (19.1%) Nocardia isolates identified, Nocardia asteroids (N. asteroids) complex (45.5%) and Nocardia brasiliensis (N. brasiliensis) (24.7%) were the most frequently isolated species, followed by Nocardia otitidiscaviarum (2.2%), Nocardiopsis dassonvillei, Actinomadura actinomadura (each 1.7%) and Nocardia transvalensis (1.1%) and also unknown spp. (23.0%). In this study, most species (54.4%) of Nocardia, especially N. asteroides complex were isolated from soils with pH: 7.01-8, whereas in pH: 8.01-9 more N. brasiliensis was isolated. The most Nocardia spp. was detected from regions with semi-nomadic and temperate climate (41.1%).

Conclusions

N. asteroids complex is more prevalent in Isfahan province and soil can be a potential source of nocardiosis infections. It is to be considering that climate and soil pH are involved in the frequency and diversity of aerobic Actinomycetes.     

An Australia-wide epidemic of Pseudomonas pickettii bacteraemia due to contaminated "sterile" water for injection

Nineteen cases of Pseudomonas pickettii bacteraemia and one case of Pseudomonas cepacia bacteraemia were identified in an Australia-wide outbreak of nosocomial sepsis associated with contaminated water for injection. The contamination was limited to one batch of commercially produced water for injection. Four different organisms were identified (three biotypes of P. pickettii and one of P. cepacia). However, P. pickettii biotype 1 appeared to be relatively more virulent than the other biotypes as it was the only identified organism in blood cultures in nearly all cases of sepsis. The ampoules of "sterile" water were each contaminated with approximately 10(3) organisms per millilitre. The lack of an Australian central reporting system for bacteraemia delayed the recognition of this outbreak.     

耐万古霉素肠球菌表型及基因型分析

目的：对耐万古霉素的肠球菌进行表型及基因型分析，了解临床耐万古霉素肠球菌(VRE)的流行状况。指导临床合理使用抗生素。方法：48株肠球菌，按常规方法进行鉴定，并采用琼脂筛选法对耐万古霉素肠球菌进一步筛选。将筛选出的3株耐万古霉素肠球菌，以多重PCR法进行基因分型。结果：共检出3株对万古霉素耐药肠球菌，其中1株为天然耐万古霉素株。基因型分析结果为1株VanA型，1株VanCl型，另1株基因型不明。结论：已发现3株VRE。临床应合理使用抗生素，以防止VRE的爆发流行。     

啮齿类螺杆菌的分离鉴定

目的对分离自啮齿类实验动物的17株螺杆菌进行鉴定,以期获得生物学、遗传学特性典型的菌株作为模式菌株。方法通过生物学特性、16S rRNA序列测定、系统发育树及超微结构分析,对螺杆菌进行种的鉴定。结果确定所分离的17株菌株分为两大类,其中一类为胆汁螺杆菌(H.bilis);另外一类属于螺杆菌属成员。结论所分离到的菌株分别是胆汁螺杆菌和未鉴定到种的螺杆菌,其中胆汁螺杆菌与ATCC51630模式菌株16SrRNA序列比较,相似性达99.6%,可作为检测用的模式菌株。     

Nocardia asteroides sinusitis. Presentation as a trimethoprim-sulfamethoxazole responsive fever of unknown origin.

A patient with a yearlong fever of unknown origin responded to trimethoprim-sulfamethoxazole and was discovered to have culture-proved Nocardia asteroides sinusitis, with absence of detectable disease in other organs. An inhalational route is postulated as the mode of entry of the organism, with localization in the maxillary sinus.     

16S rDNA序列分析在临床不常见细菌鉴定中的初步应用

目的建立细菌16S rDNA序列分析技术并探讨其在临床不常见细菌鉴定中的应用价值。方法收集经全自动微生物分析系统鉴定其可靠率在99%及以上的常见细菌10株和常规方法难以鉴定或少见菌17株,通过PCR扩增16S rDNA v3-v4区基因片段并进行序列测定,序列与16SpathDB 2.0数据库上已知细菌的16S rDNA序列进行比对分析确定细菌种属。当v3-v4序列片段不能明确鉴定细菌时,结合16S rDNA长片段测序及管家基因dnaJ和rpoB的序列做进一步分析。结果经16S rDNA v3-v4区序列分析,10株(100%)临床常见菌都能鉴定到种水平,其可靠性大于98.0%。17株不常见菌株中,10株(58.8%)细菌只与1种菌相似性大于98.0%,成功鉴定到单个种水平;6株(35.3%)细菌与不止1种菌的相似比大于98.0%;1株菌与已知菌相似性在96.0%~98.0%之间只能鉴定到属(棒状杆菌)。进一步结合16S rDNA长片段及管家基因dnaJ和rpoB的序列分析,所有菌株均可鉴定到单个种水平。结论结合16S rDNA和管家基因序列分析,可用于临床常见及不常见细菌的分子鉴定。     

MALDI-TOF MS技术在鉴定临床少见非发酵菌中的应用

摘要：目的  评价基质辅助激光解析电离飞行时间质谱（MALDI-TOF MS）技术在快速鉴定临床少见非发酵菌中的应用价值。方法  选取2013年7月-2016年10月安徽医科大学第一附属医院检验科分离的67株临床少见非发酵菌,包括痰液分离的21株,尿液分离的13株,血液分离的9株,骨髓液分离的9株,分泌物分离的8株,腹透液分离的2株,脑脊液分离的1株,胆汁分离的1株,其他体液分离的3株。以16S核糖体RNA（16SrRNA）序列分析为金标准,比较MALDI-TOF MS与Vitek2 Compact鉴定系统鉴定的准确性。结果  67株临床少见非发酵菌中,MALDI-TOF MS将66株（98.5%）鉴定到种水平,1株（1.5%）鉴定到属水平;Vitek2 Compact鉴定系统将59株（88.1%）鉴定到种水平,8株（11.9%）未能鉴定;67株少见非发酵菌均扩增到16SrRNA目的基因片段并成功测序,67株（100.0%）都成功鉴定到种水平。以16SrRNA序列分析为金标准,MALDI- TOF MS和Vitek2 Compact鉴定系统正确鉴定到种水平准确性分别为98.5%（66/67）和88.1%（59/67）。结论  与传统的细菌生化反应方法相比,MALDI-TOF MS技术操作简便、耗时短、费用低,鉴定结果与16SrRNA序列分析的符合率高,可以提高临床对少见非发酵菌鉴定的准确率。

     

应用巢氏PCR-RFLP直接检测结核病临床标本中结核分支杆菌链霉素耐药基因型

目的：了解结核分支杆菌链霉素耐药基因突变情况,建立快速检测耐药突变株的方法。方法：通过ＰＣＲ－ＳＳＣＰ、ＰＣＲ－ＲＦＬＰ和巢氏ＰＣＲ－ＲＦＬＰ分析４０株结核分支杆菌临床分离株和１５例临床标本的ｒｐｓＬ基因突变的部位和性质。结果：１０株药物敏感株的ｒｐｓＬ双链泳动正常、并可被ＭｂｏⅡ消化;３０株耐ＳＭ分离株中,２５株双链泳动异常,２４株不被消化;１５例临床标本中,常规ＰＣＲ扩增７例ｒｐｓＬ阳性,巢氏ＰＣＲ１５例均阳性,４例ｒｐｓＬ双链泳动异常、并不被ＭｂｏⅡ消化。结论：通过常规ＰＣＲ－ＳＳＣＰ和ＰＣＲ－ＲＦＬＰ可快速检测结核分支杆菌耐ＳＭ分离株４３位密码子点突变,而通过巢氏ＰＣＲ－ＲＦＬＰ可直接检测大多数临床标本中结核分支杆菌ＳＭ耐药基因型。     

甄氏外瓶霉的核糖体基因分型研究

目的:应用核糖体基因(rDNA)序列对甄氏外瓶霉进行分型研究.方法:试验菌株包括甄氏外瓶霉及其变种20株(模式株、标准株、临床及自然分离株),丛梗孢外瓶霉8株,威尼克外瓶霉1株(标准株及临床和环境分离株),疣状瓶霉1株.煮沸冷冻法提取DNA,常规PCR扩增核糖体基因及其转录间隔区,应用DNA多序列分析等方法进行研究.结果:甄氏外瓶霉核糖体基因及其转录间隔区变异较大,DNA序列分析显示,不同来源、不同致病性的甄氏外瓶霉可分为7型,个别菌种与其他甄氏外瓶霉的遗传距离较大.结论:甄氏外瓶霉核糖体基因及其转录间隔区序列变异较大,个别菌株的归属有待于进一步研究.     

Differentiation of Helicobacter pylori isolates by polymerase chain reaction—restriction fragment length polymorphism

Objective: To investigate the association between the diversity of urease gene and urease activity of clinical isolates of Helicobacter pylori (H. pylori). Methods: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of urease gene and rapid urease activity test were used to study the urease activity of different clinical isolates of H. pylori. Results: H. pylori clinical isolates were divided into 4 types according to their PCR-RFLP results of urease gene and urease activity. Type Ⅰ , possessing strong urease activity (0. 11) and presented 1 fragment of 1. 7 kb by PCR-RFLP, had close relations with gastric ulcer; type Ⅱ , with the weakest urease activity (0. 07) and 2 fragments (1. 3 and 0. 4 kb respectively) , was associated with duodenal bulb ulcer; type Ⅲ , with the strongest urease activity (0. 12) and 2 fragments (0. 4 and 0. 17 kb) with or without 1 fragment (0. 23 or 0. 37 kb) , was responsible for gastritis; type Ⅳ , with weak urease activity (0. 09) and 2 fragments     

The inherent characteristics and DNA polymorphism of Vibrio cholerae and other vibrios

Objective To investigate the inherent characteristics of Vibrio cholerae (V.cholerae) and other vibrios and their relationship.Methods Polymerase chain reaction (PCR), DNA sequence analysis, randomly amplified polymorphic DNA (RAPD) analysis and average linkage cluster analysis were used to study 3 isolates of V.cholerae strains O139, three isolates O1 biot ype El Tor, four isolates O1 biotype classical and 3 other vibrios.Results V.cholerae O139 contained the genomic sequences of ctx A2- B as well as V.cholerae O1.V.cholerae and others vibrios were divided into 4 groups by fingerprint patterns of RAPD, that is ① V.cholerae O139 and V.cholerae O1 El Tor; ② V.cholerae O1 classical; ③ V.para heamolyticus and V.vulnificus and ④ V.flluvialis.V.cholerae O139 DNA fingerprint of RAPD was consistent with the El Tor biotype: average linkage cluster distance was 0, and slightly different from the classical b iotype, with a distance of 2.07.It was much more different from vibrio par aheamolyticus and others, with a distance of 6.76-8.54Conclusion V.cholerae and other vibrios are polymorphic in inherent characteristics. The inherent characteristics of V.cholerae O139 are the same as El Tor b iotype.O139 may have evolved from the El Tor biotype.The inherent char acteristics of vibrio paraheamolyticus are the same as vibrio vulnificus.     

从马拉色菌毛囊炎患者皮损和非皮损区分离和鉴定马拉色菌

目的研究马拉色菌毛囊炎患者皮损区和非皮损区马拉色菌菌种的检出以及菌种的构成。方法采用菜子油培养基,根据菌落形态和生理生化特点进行菌种鉴定。结果从毛囊内共分离出326株马拉色菌,其中球形马拉色菌255株,占78.22%;合轴马拉色菌54株,占16.56%;糠秕马拉色菌17株,占5.22%。从皮损部位疹间皮肤皮屑分离马拉色菌298株,其中合轴马拉色菌221株,占74.16%;糠秕马拉色菌43株,占14.43%;球形马拉色菌30株,占10.07%;钝形马拉色菌4株,占1.34%。菌种检出率和菌种构成差异均有显著性(P<0.05和P<0.0001)。结论球形马拉色菌是马拉色菌毛囊炎的优势致病菌,正常皮肤则以合轴马拉色菌为主。