16s rRNA sequencing

16S rRNA 测序
  • 文章类型: Case Reports
    背景:硝基还原假单胞菌是一种非发酵,革兰氏阴性,通常居住在土壤中的杆状细菌,特别是被油盐水污染的土壤。据我们所知,以前没有人感染硝基还原丙酸杆菌的病例报道。这里,我们介绍了第一例有记载的菌血症患者中由硝基还原丙酸杆菌引起的胆管炎病例。
    方法:一名患有晚期胰腺神经内分泌肿瘤的46岁日本男子因发热和寒战住院。入院前四天,患者出现右上腹痛。两天后,他还发烧和发冷。入院当天进行内镜逆行胰胆管造影术,患者被诊断为患有与支架功能障碍相关的胆管炎。从血液培养物中分离出革兰氏阴性棒,但尝试使用VITEK2和基质辅助激光解吸/电离飞行时间质谱(MALDI-TOFMS)与VITEKMSver鉴定细菌。4.7.1(BioMérieuxJapanCo.Ltd.,东京,日本)没有成功。最后,使用MALDI-TOFMS和MALDIBiotyper(BrukerDaltonicsCo.,Ltd.,Billerica,MA,美国)和16S核糖体RNA测序。尽管对病人进行了彻底的采访,他否认接触受污染的土壤。根据药敏结果,患者静脉注射头孢吡肟和口服环丙沙星治疗16天,取得良好的治疗效果。在第28天的门诊随访中,患者的一般情况良好。
    结论:这是首例报道的由硝基还原丙酸杆菌引起的血流感染的人类胆管炎病例。本报告为临床医生提供了准确诊断硝基还原丙酸杆菌的临床表现和诊断方法的新见解。以及治疗指导。
    BACKGROUND: Pseudomonas nitroreducens is a non-fermenting, gram-negative, rod-shaped bacterium commonly inhabiting soil, particularly soil contaminated with oil brine. To our knowledge, no cases of human infection with P. nitroreducens have been previously reported. Here, we present the first documented case of cholangitis caused by P. nitroreducens in a patient with bacteremia.
    METHODS: A 46-year-old Japanese man with an advanced pancreatic neuroendocrine tumor was hospitalized with fever and chills. Four days before admission, the patient developed right upper abdominal pain. Two days later, he also experienced fever and chills. Endoscopic retrograde cholangiopancreatography was performed on the day of admission, and the patient was diagnosed as having cholangitis associated with stent dysfunction. Gram-negative rods were isolated from blood cultures, but attempts to identify the bacteria using VITEK2 and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with VITEK MS ver. 4.7.1 (bioMérieux Japan Co. Ltd., Tokyo, Japan) were unsuccessful. Finally, the organism was identified as P. nitroreducens using MALDI-TOF MS with a MALDI Biotyper (Bruker Daltonics Co., Ltd., Billerica, MA, USA) and 16 S ribosomal RNA sequencing. Despite thorough interviews with the patient, he denied any exposure to contaminated soil. The patient was treated with intravenous cefepime and oral ciprofloxacin for 16 days based on susceptibility results, achieving a good therapeutic outcome. At the outpatient follow-up on day 28, the patient was in good general condition.
    CONCLUSIONS: This is the first reported human case of cholangitis with bloodstream infection caused by P. nitroreducens. This report provides clinicians with novel insights into the clinical manifestations and diagnostic methods necessary for the accurate diagnosis of P. nitroreducens, along with guidance on treatment.
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  • 文章类型: Journal Article
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  • 文章类型: Case Reports
    食肉动物是人类感染的一种极为罕见的病原体。该病例报告了食蟹猴菌血症感染,如果使用不适当的诊断方法,极有可能导致误诊。
    一名没有基础疾病的29岁中国男性患者因车祸受伤两次住院。在第二次住院期间,进行了腹部手术,手术后出现高烧。从血液中分离出一种O.splanchnicus菌株,并通过MALDI-TOF-MS和16SrRNA基因分析进行确认。最后,患者通过使用抗生素成功康复,补液和白蛋白输入。
    这是中国首例O.splanchnicus菌血症。我们简要回顾了有关人类O.splanchnicus感染的病例。O.斯普兰尼克,作为正常肠道菌群的一部分,以其抗肿瘤和免疫调节特性而闻名,它很少从临床样本中分离出来。此案例说明了O.splanchnicus作为病原体的潜力,并建议注意使用新的和先进的方法,例如MALDI-TOFMS和16SrRNA基因测序,以从临床样品中鉴定很少分离的物种。
    UNASSIGNED: Odoribacter splanchnicus is an extremely rare pathogen of human infection. This case reports bacteremia infection of O. splanchnicus, which is highly likely to result in misdiagnosis if inappropriate diagnostic method are used.
    UNASSIGNED: A 29-year-old Chinese male patient with no underlying disease was hospitalized twice for injuries caused by a car accident. During the second hospitalization, abdominal surgery was performed and high fever developed after the surgery. A strain of O. splanchnicus was isolated from the blood and confirmed by MALDI-TOF-MS and 16S rRNA gene analysis. Finally, the patient recovered successfully by using antibiotics, fluid replacement and albumin input.
    UNASSIGNED: This is the first case of O. splanchnicus bacteremia in China. We present a brief review of the cases concerning O. splanchnicus infection in humans. O. splanchnicus, as part of the normal intestinal flora, is well known for its anti-tumor and immune regulating properties, it is rarely isolated from clinical samples. This case illustrates the potential of O. splanchnicus as a pathogen and suggests attention to the use of new and advanced methods like MALDI-TOF MS and 16S rRNA gene sequencing to identify rarely isolated species from clinical samples.
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  • 文章类型: Journal Article
    口腔菌群与各种免疫相关疾病有关。本文探讨了寻常型天疱疮(PV)患者口腔菌群的特点,并分析了口腔菌群与PV的相关性。
    本病例对照研究包括22名未经治疗的PV患者和12名健康对照(HC)。使用16SrRNAIlluminaMiSeq方法通过高通量测序评估唾液微生物组的特征,并确定PV组和HC组之间的差异。利用京都基因和基因组百科全书(KEGG)数据库筛选关键的代谢途径,并初步探索PV发生和发展的潜在机制。
    PV组口腔菌群丰度明显低于HC组,并且有特征性的变化。PV组的Prevotella和农杆菌的相对丰度显著高于HC组(P<0.05)和奈瑟菌,Lautropia,梭菌显著降低(P<0.05)。PV中Prevotella与血清Dsg3水平呈线性相关。KEGG通路分析表明,PV组和HC组之间的9种代谢通路存在显着差异(P<0.05)。即碳水化合物代谢,消化系统,神经退行性疾病,聚糖生物合成和代谢,耐药性:抗菌药物,传染病:病毒性,循环系统,排泄系统,和神经系统。
    PV患者口腔菌群呈现特征性变化,几种代谢途径受到影响,包括N-聚糖生物合成和代谢。普雷沃氏菌属。在PV中似乎最需要关注。我们认为口腔菌群失调有助于PV的发生和发展。
    UNASSIGNED: Oral flora is related to various immune-related diseases. Herein we explored the characteristics of oral flora in patients with pemphigus vulgaris (PV) and analyzed the correlation between oral flora and PV.
    UNASSIGNED: Twenty-two untreated patients with PV and 12 healthy controls (HC) were included in this case-control study. The characteristics of salivary microbiome were assessed by high-throughput sequencing using the 16S rRNA Illumina MiSeq approach, and differences between the PV and HC groups were determined. The Kyoto Encyclopedia of Genes and Genomes (KEGG) database was applied to screen key metabolic pathways and preliminarily explore potential mechanisms underlying PV occurrence and development.
    UNASSIGNED: The abundance of oral flora in the PV group was significantly lower than that in the HC group, and there were characteristic changes. The relative abundance of Prevotella and Agrobacterium in the PV group was significantly higher than that in the HC group (P < 0.05) and that of Neisseria, Lautropia, and Fusobacterium was significantly lower (P < 0.05). There was a linear correlation between Prevotella and serum Dsg3 level in PV. KEGG pathway analyses indicated significant differences in nine metabolic pathways between the PV and HC groups (P < 0.05), namely carbohydrate metabolism, digestive system, neurodegenerative disease, glycan biosynthesis and metabolism, drug resistance: antimicrobial, infectious disease: viral, circulatory system, excretory system, and nervous system.
    UNASSIGNED: The oral flora of patients with PV presented characteristic changes, and several metabolic pathways were affected, including N-glycan biosynthesis and metabolism. Prevotella spp. appear to require the most attention in PV. We believe that oral flora dysbacteriosis contributes to PV occurrence and development.
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    文章类型: Review
    拟白喉棒状杆菌,属于口咽微生物群的革兰氏阳性棒,通常描述为肺部感染,尤其是免疫功能低下的患者。本文描述了一例罕见的天然主动脉感染性心内膜炎(IE),并回顾了有关类似病例的文献。一名62岁的男子从小就患有风湿热,因大植被(15.8X8.3mm)的白喉梭菌引起的高热IE手术治疗。MALDI-TOF-MS来自阳性血液培养物中分离的菌株,鉴定为假白喉梭菌(2.34),和来自瓣膜样品的16SrRNA测序证实了鉴定。对25例病例的总结表明,假性白喉梭菌引起的IE的结局不好。文献综述表明,在心血管背景下的血液培养物中发现的这种药物值得仔细研究,因为预后不良经常发生。
    Corynebacterium pseudodiphtheriticum, a Gram-positive rod belonging the oropharynx microbiota, is usually described in pulmonary infections, especially in immunocompromised patients. This paper describes a rare case of native aortic infectious endocarditis (IE) and reviews the literature on similar cases. A 62-year-old man with rheumatic fever since childhood was hospitalized for surgical treatment of a febrile IE due to C. diphtheriticum with a large vegetation (15.8 X 8.3 mm). MALDI-TOF-MS from strain isolated in positive blood cultures identified C. pseudodiphtheriticum (2.34), and 16S rRNA sequencing from the valve sample confirmed the identification. The summary of 25 cases shows that the outcome of an IE due to C. pseudodiphtheriticum is bad. The review of the literature shows that this agent found in blood cultures in a cardiovascular context deserves to be explored meticulously because an unfavorable prognosis is frequent.
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  • 文章类型: Journal Article
    目的:关于肠道菌群如何改变特发性帕金森病(PD)脑肠轴的研究引起了越来越多的兴趣。我们旨在确定PD患者的肠道微生物群是否发生改变,以及PD的非运动症状和疾病持续时间是否与患者中微生物群的改变有关。
    方法:使用16SrRNA扩增子测序分析了从84名受试者(42-PD患者和42名健康配偶)获得的粪便样品中的微生物分类群。
    结果:我们观察到在门水平上Firmicutes的显着减少和Verrucoomicota的显着增加。在家庭层面,与健康配偶相比,PD患者的乳杆菌和Akkermansiaceae显着增加,而CoriobacterialesIncertaeSedis显着减少。属水平比较推断丰度仅在乳杆菌属中显著增加,而落叶草科ND3007组的丰度,Tyzzerella,镰刀菌,嗜真菌组和隔黄流球菌组均下降。我们确定普雷沃氏菌属的丰度下降,但在PD患者中并不明显。此外,我们发现有和没有非运动症状的患者的微生物群组成存在差异.
    结论:我们观察到PD患者与其健康配偶的肠道菌群组成存在差异。我们的研究结果表明,疾病持续时间影响微生物群组成,进而影响PD非运动症状的发展。这项研究是土耳其PD患者肠道微生物群研究以及微生物群对PD非运动症状的可能影响方面的第一项。
    OBJECTIVE: The investigations related to how gut microbiota changes the brain-gut axis in idiopathic Parkinson\'s disease (PD) attract growing interest. We aimed to determine whether gut microbiota is altered in PD patients and whether non-motor symptoms of PD and disease duration had any relation with alterations of microbiota profiles among patients.
    METHODS: Microbial taxa in stool samples obtained from 84 subjects (42-PD patients and 42-healthy spouses) were analyzed using 16S rRNA amplicon-sequencing.
    RESULTS: We observed a significant decrease of Firmicutes and a significant increase of Verrucomicrobiota at the phylum level. At the family level, Lactobacillaceae and Akkermansiaceae were significantly increased and Coriobacteriales Incertae Sedis were significantly decreased in the PD patients compared to their healthy spouses. Genus level comparison inferred significant increase in abundance only in Lactobacillus while the abundance of Lachnospiraceae ND3007 group, Tyzzerella, Fusicatenibacter, Eubacterium hallii group and Ruminococcus gauvreauii group were all decreased. We determined that the abundance of Prevotella genus decreased, but not significantly in PD patients. In addition, we found differences in microbiota composition between patients with and without non-motor symptoms.
    CONCLUSIONS: We observed differences in gut microbiota composition between PD patients and their healthy spouses. Our findings suggest that disease duration influenced microbiota composition, which in turn influenced development of non-motor symptoms in PD. This study is the first in terms of both gut microbiota research in Turkish PD patients and the probable effect of microbiota on non-motor symptoms of PD.
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  • 文章类型: Journal Article
    与人类活动相关的重金属污染在热带海湾备受关注。微生物可能对重金属高度敏感。尽管如此,关于重金属对热带海湾沉积物中微生物结构的影响知之甚少。在这项研究中,采用16SrRNA基因测序和潜在生态风险指数分析对9种金属(砷、铅,镉,钴,铬,铜,锌,锰,和镍)和巴门湾沉积物中的细菌群落,中国。我们的结果表明,巴门湾具有相当大的生态风险,镉具有最高的单项潜在生态风险。此外,单个金属污染与细菌群落组成相关,但与细菌α多样性无关。砷是对细菌群落结构影响最大的金属。我们的发现为热带海湾重金属污染的监测和修复提供了新的见解。
    Heavy metal pollution associated with human activity is of big concern in tropical bays. Microorganisms may be highly sensitive to heavy metals. Nonetheless, little is known about effects of heavy metals on microbial structure in tropical bay sediments. In this study, 16S rRNA gene sequencing and potential ecological risk index analysis were used to analyze the relationships between nine metals (arsenic, lead, cadmium, cobalt, chromium, copper, zinc, manganese, and nickel) and bacterial communities in the sediments of Bamen Bay, China. Our results showed that Bamen Bay was under a considerable ecological risk and cadmium had the highest monomial potential ecological risk. In addition, individual metal contamination correlated with bacterial community composition but not with bacterial α-diversity. Arsenic was the metal influencing bacterial community structure the most. Our findings provide a novel insight into the monitoring and remediation of heavy metal pollution in tropical bays.
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  • 文章类型: Journal Article
    背景:许多奶牛在从妊娠晚期过渡到泌乳早期时经历能量不足的状态。这项研究的目的是1)确定泌乳早期奶牛的肠道微生物组是否表明高酮血症的发展,和2)识别可能告知健康状况的微生物特征。我们进行了一项前瞻性巢式病例对照研究,其中奶牛在产牛前14至7天被招募,并在牛奶中(DIM)的前14天被招募。高酮症奶牛(HYK,n=10)基于其第一个14DIM内的血液β-羟基丁酸酯(BHB)浓度1.2mmol/L进行分类。对于每一头HYK母牛,两个非HYK(CON,n=20)母牛通过产次和3个DIM进行匹配,BHB<1.2mmol/L每日血液BHB测量用于确认CON母牛保持健康状态;一些CON母牛在匹配后显示BHB1.2mmol/L,这些母牛被重新分类为对照-HYK(C-HYK,n=9)。在诊断或匹配当天收集瘤胃和粪便样品并进行16SrRNA分析。
    结果:分类群丰度没有差异,或者阿尔法和贝塔多样性,在CON中观察到,C-HYK,和HYK健康小组的粪便微生物组。在所有健康状况下,基于β多样性分析检测到相似的微生物组组成,然而,发现CON和HYK奶牛的瘤胃微生物组存在显着差异。有趣的是,在C-HYK奶牛瘤胃和粪便微生物组中观察到高度相似的微生物组组成,这表明,这些最初看起来健康,迟发性高酮血症的动物个体彼此高度相似。与CON和HYK组相比,这些C-HYK母牛在其瘤胃微生物组中具有显著较低丰度的反刍动物球菌2。用于计算微生物丰度相对于健康状况的对数倍数变化的多项回归未发现具有预测价值,因此,对于确定某些微生物特征在预测健康状况中的作用没有帮助。
    结论:在C-HYK奶牛瘤胃中观察到较低的Ruminococus2相对丰度,这表明这些奶牛可能在降解纤维素方面效率较低,尽管Ruminococusspp的机制作用。在瘤胃中的新陈代谢还没有完全了解。没有观察到经历不同能量短缺水平的奶牛的粪便或瘤胃微生物组的实质性差异,表明高酮血症可能不会受到肠道微生物组成的很大影响,反之亦然。需要使用更高分辨率的组学方法如元转录组学或元蛋白质组学的进一步研究来破译确切的机制。
    BACKGROUND: Many dairy cows experience a state of energy deficit as they transition from late gestation to early lactation. The aims of this study were to 1) determine if the development of hyperketonemia in early lactation dairy cows is indicated by their gut microbiome, and 2) to identify microbial features which may inform health status. We conducted a prospective nested case-control study in which cows were enrolled 14 to 7 days before calving and followed through their first 14 days in milk (DIM). Hyperketonemic cows (HYK, n = 10) were classified based on a blood β-hydroxybutyrate (BHB) concentration 1.2 mmol/L within their first 14 DIM. For each HYK cow, two non-HYK (CON, n = 20) cows were matched by parity and 3 DIM, with BHB < 1.2 mmol/L. Daily blood BHB measures were used to confirm CON cows maintained their healthy status; some CON cows displayed BHB 1.2 mmol/L after matching and these cows were reclassified as control-HYK (C-HYK, n = 9). Rumen and fecal samples were collected on the day of diagnosis or matching and subjected to 16S rRNA profiling.
    RESULTS: No differences in taxa abundance, or alpha and beta diversity, were observed among CON, C-HYK, and HYK health groups for fecal microbiomes. Similar microbiome composition based on beta diversity analysis was detected for all health statuses, however the rumen microbiome of CON and HYK cows were found to be significantly different. Interestingly, highly similar microbiome composition was observed among C-HYK cow rumen and fecal microbiomes, suggesting that these individual animals which initially appear healthy with late onset of hyperketonemia were highly similar to each other. These C-HYK cows had significantly lower abundance of Ruminococcus 2 in their rumen microbiome compared to CON and HYK groups. Multinomial regressions used to compute log-fold changes in microbial abundance relative to health status were not found to have predictive value, therefore were not useful to identify the role of certain microbial features in predicting health status.
    CONCLUSIONS: Lower relative abundance of Ruminococcus 2 in C-HYK cow rumens was observed, suggesting these cows may be less efficient at degrading cellulose although the mechanistic role of Ruminococcus spp. in rumen metabolism is not completely understood. Substantial differences in fecal or rumen microbiomes among cows experiencing different levels of energy deficit were not observed, suggesting that hyperketonemia may not be greatly influenced by gut microbial composition, and vice versa. Further studies using higher resolution -omics approaches like meta-transcriptomics or meta-proteomics are needed to decipher the exact mechanisms at play.
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  • 文章类型: Case Reports
    药用水蛭(Hirudomedicinalis)以手术和非手术方式使用。Morganellamorganii是一种机会性和人畜共患的致病菌,可引起严重的临床并发症。在这项研究中,我们与世隔绝,发现并鉴定了M.morganii感染的H.medicinalis。我们在所有发炎和肿胀的Hirudomedicinalis样本中检测并鉴定了morganii。分离物的16SrRNA序列证实了M.morganii的所有菌株。所有菌株均对头孢曲松敏感,头孢噻夫,丹氧氟沙星,环丙沙星,恩诺沙星,土霉素,和美罗培南对红霉素耐药,阿莫西林,氨苄青霉素,头孢唑啉,粘菌素,青霉素G,还有林可霉素.这种致病菌是一种人畜共患病原体,监测这种细菌的患病率对于用于人类医疗和护理的水蛭来说是非常必要的。最后,在本病例报告研究中,所有感染的水蛭均得到成功治疗.
    Medicinal leeches (Hirudo medicinalis) are used in surgical and non-surgical manners. Morganella morganii is an opportunistic and zoonotic pathogenic bacterium causing serious clinical complications. In this study, we isolated, discovered and characterized M. morganii-infected H. medicinalis. We detected and identified M. morganii in all inflamed and swollen Hirudo medicinalis samples. The 16S rRNA sequence of the isolates confirmed all strains of M. morganii. All strains were sensitive to Ceftriaxone, Ceftiofur, Danofloxacin, Ciprofloxacin, Enrofloxacin, Oxytetracycline, and Meropenem and were resistant to Erythromycin, Amoxicillin, Ampicillin, Cefazolin, Colistin, Penicillin G, and Lincomycin. This pathogenic bacterium is a zoonotic pathogen, and monitoring the prevalence rate of this bacteria is strongly necessary for leeches used in human medical treatment and care. Finally, all infected leeches were treated successfully in this case report study.
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  • 文章类型: Case Reports
    Rosorum是革兰氏阴性兼性厌氧菌,2011年从巴斯德科重新分类。已在哺乳动物的胃肠道和呼吸道中检测到;然而,人类感染的报告很少。我们报告了一例腹部脓肿,其中检测到N.rosorum;通过引流和抗菌治疗成功治疗。常规实验室测试,例如基质辅助激光解吸/电离飞行时间质谱和使用生化表型的识别系统,无法识别N.rosorum。相反,它被误认为是其他巴氏杆菌科物种,包括Aggregatibacterspp.或者巴斯德菌。需要对16SrRNA进行测序以鉴定罗索酵母。我们建议应用简单的方法,如吲哚生产,氧化酶,和过氧化氢酶测试,区分N.rosorum从基因相似的物种。
    Necropsobacter rosorum is a gram-negative facultative anaerobe, which was reclassified from the family Pasteurellaceae in 2011. It has been detected in the gastrointestinal and respiratory tracts of mammals; however, reports of infection in humans are scarce. We report a case of an abdominal abscess in which N. rosorum was detected; it was successfully treated with drainage and antimicrobial therapy. Routine laboratory testing such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and an identification system using biochemical phenotypes could not identify N. rosorum. Instead, it was misidentified as other Pasteurellaceae species, including Aggregatibacter spp. or Pasteurella spp. Sequencing of 16S rRNA was required to identify N. rosorum. We suggest the application of simple methods, such as indole production, oxidase, and catalase tests, to differentiate N. rosorum from genetically similar species.
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