metagenomic sequencing

宏基因组测序
  • 文章类型: Journal Article
    植物圈细菌可以调节影响海洋生态系统的不同藻华的动态。球形假囊藻可以在孤立的自由活细胞和集落之间交替,后者的形态类型在开花期间占主导地位。这些水华形成的潜在机制受到了很多关注。高通量测序结果表明,菌落和孤立菌株之间的细菌群落组成在细菌组成和功能上存在显着差异。发现SM1A02和Haliea属仅在菌落菌株中检测到,并有助于菌落中的铵积累,在生产DMS的优良菌落菌株中,硫杆菌属丰富。此外,两个菌落菌株的细菌群落表现出较强的碳和硫代谢能力,能量代谢,维生素B合成,和信号转导,提供无机和有机营养,促进与宿主藻类的紧密交流,从而促进生长和开花发展。
    Phycosphere bacteria can regulate the dynamics of different algal blooms that impact marine ecosystems. Phaeocystis globosa can alternate between solitary free-living cells and colonies and the latter morphotype is dominate during blooms. The mechanisms underlying the formation of these blooms have received much attention. High throughput sequencing results showed that the bacterial community composition differed significantly between colony and solitary strains in bacterial composition and function. It was found that the genera SM1A02 and Haliea were detected only among the colony strains and contribute to ammonium accumulation in colonies, and the genus Sulfitobacter was abundant among the colony strains that were excellent at producing DMS. In addition, the bacterial communities of the two colony strains exhibited stronger abilities for carbon and sulfur metabolism, energy metabolism, vitamin B synthesis, and signal transduction, providing inorganic and organic nutrients and facilitating tight communication with the host algae, thereby promoting growth and bloom development.
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  • 文章类型: Journal Article
    背景:幽门螺杆菌(H.幽门螺杆菌)感染与胃肠道疾病密切相关。我们的初步研究表明,幽门螺杆菌感染对胃溃疡(GU)或十二指肠溃疡(DU)患者的粘膜微生物组结构有重大影响。
    目的:探讨幽门螺杆菌感染和黏膜微生物群对溃疡性疾病发病及进展的影响。
    方法:幽门螺杆菌感染和GU或DU的患者,和没有幽门螺杆菌感染的健康个体被包括在内。获得胃或十二指肠粘膜样品并进行宏基因组测序。分析了微生物群落的组成及其在粘膜组织中的代谢功能。
    结果:与健康个体相比,幽门螺杆菌阳性GU患者的胃粘膜微生物群以幽门螺杆菌为主,生物多样性显著减少。组间微分函数,富含幽门螺杆菌阳性GU患者,都来自幽门螺杆菌,特别是那些涉及转移RNA的糖苷修饰和去甲基甲基甲萘醌或甲萘醌的合成。在合成途径中检测到uibE基因的显著富集。幽门螺杆菌阳性DU患者和健康对照组之间的微生物多样性没有显着差异。
    结论:H.幽门螺杆菌感染显著改变了胃微生物群结构,多样性,和生物学功能,这可能是GU的重要促成因素。
    BACKGROUND: Helicobacter pylori (H. pylori) infection is closely associated with gastrointestinal diseases. Our preliminary studies have indicated that H. pylori infection had a significant impact on the mucosal microbiome structure in patients with gastric ulcer (GU) or duodenal ulcer (DU).
    OBJECTIVE: To investigate the contributions of H. pylori infection and the mucosal microbiome to the pathogenesis and progression of ulcerative diseases.
    METHODS: Patients with H. pylori infection and either GU or DU, and healthy individuals without H. pylori infection were included. Gastric or duodenal mucosal samples was obtained and subjected to metagenomic sequencing. The compositions of the microbial communities and their metabolic functions in the mucosal tissues were analyzed.
    RESULTS: Compared with that in the healthy individuals, the gastric mucosal microbiota in the H. pylori-positive patients with GU was dominated by H. pylori, with significantly reduced biodiversity. The intergroup differential functions, which were enriched in the H. pylori-positive GU patients, were all derived from H. pylori, particularly those concerning transfer RNA queuosine-modification and the synthesis of demethylmenaquinones or menaquinones. A significant enrichment of the uibE gene was detected in the synthesis pathway. There was no significant difference in microbial diversity between the H. pylori-positive DU patients and healthy controls.
    CONCLUSIONS: H. pylori infection significantly alters the gastric microbiota structure, diversity, and biological functions, which may be important contributing factors for GU.
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  • 文章类型: Journal Article
    妊娠期糖尿病(GDM)对母婴微生物组轨迹的影响仍然知之甚少。利用264个母婴二元的大规模纵向粪便样本,我们介绍了母亲在整个怀孕期间和婴儿出生后第一年的肠道微生物组轨迹.GDM母亲在妊娠期具有不同的微生物组多样性和组成。GDM在婴儿的肠道微生物组上留下指纹,被交付模式所混淆。Further,梭菌属物种与雄性后代12个月时较大的头围呈正相关,而雌性则不相关。有男性胎儿的GDM母亲的肠道微生物组显示出耗尽的肠脑模块,包括乙酸合成I和降解和谷氨酸合成II。GDM母亲的女性婴儿的肠道微生物组具有较高的组胺降解和多巴胺降解。一起,我们的综合分析表明GDM会影响母婴肠道成分,这与性二态婴儿头部发育有关。
    The impact of gestational diabetes mellitus (GDM) on maternal or infant microbiome trajectory remains poorly understood. Utilizing large-scale longitudinal fecal samples from 264 mother-baby dyads, we present the gut microbiome trajectory of the mothers throughout pregnancy and infants during the first year of life. GDM mothers had a distinct microbiome diversity and composition during the gestation period. GDM leaves fingerprints on the infant\'s gut microbiome, which are confounded by delivery mode. Further, Clostridium species positively correlate with a larger head circumference at month 12 in male offspring but not females. The gut microbiome of GDM mothers with male fetuses displays depleted gut-brain modules, including acetate synthesis I and degradation and glutamate synthesis II. The gut microbiome of female infants of GDM mothers has higher histamine degradation and dopamine degradation. Together, our integrative analysis indicates that GDM affects maternal and infant gut composition, which is associated with sexually dimorphic infant head growth.
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  • 文章类型: Journal Article
    近年来,大多数关于肠道微生物组的研究主要集中在粪便样本上,使肠粘膜中的微生物群落相对未被探索。为了解决这个差距,我们的研究使用鸟枪宏基因组学分析了20例结肠息肉患者的正常直肠粘膜和匹配粪便中的微生物组成。我们的发现揭示了这两个样本集之间微生物群落的明显区别。与粪便相比,粘膜微生物组包含较少的属,伯克霍尔德菌是粪便和粘膜之间最有区别的属,强调其对粘膜的显著影响。此外,基于微生物分类和KEGG骨科(KO)成果注解,我们探讨了直肠粘膜微生物群与年龄等因素之间的关系,性别,BMI,和息肉风险水平。值得注意的是,我们确定了这些表型的新生物标志物,例如在年龄上有拉氏梭状芽胞杆菌和阴沟肠杆菌。粘膜微生物群显示与糖转运和短链脂肪酸代谢相关的KO途径的富集。我们的综合方法不仅弥合了有关直肠粘膜微生物群落的知识鸿沟,而且还强调了人体肠道内微生物相互作用的复杂性和特异性。特别是在中国人口中。
    目的:本研究提供了具有结直肠癌风险的样本中粪便和直肠粘膜微生物群落之间差异的系统水平图。它揭示了直肠粘膜独特的微生态特征及其对健康的潜在影响。此外,它提供了对肠道微生物组在结直肠癌发病机制中的作用的新见解,并为开发新的预防和治疗策略铺平了道路。
    In recent years, most studies on the gut microbiome have primarily focused on feces samples, leaving the microbial communities in the intestinal mucosa relatively unexplored. To address this gap, our study employed shotgun metagenomics to analyze the microbial compositions in normal rectal mucosa and matched feces from 20 patients with colonic polyps. Our findings revealed a pronounced distinction of the microbial communities between these two sample sets. Compared with feces, the mucosal microbiome contains fewer genera, with Burkholderia being the most discriminating genus between feces and mucosa, highlighting its significant influence on the mucosa. Furthermore, based on the microbial classification and KEGG Orthology (KO) annotation results, we explored the association between rectal mucosal microbiota and factors such as age, gender, BMI, and polyp risk level. Notably, we identified novel biomarkers for these phenotypes, such as Clostridium ramosum and Enterobacter cloacae in age. The mucosal microbiota showed an enrichment of KO pathways related to sugar transport and short chain fatty acid metabolism. Our comprehensive approach not only bridges the knowledge gap regarding the microbial community in the rectal mucosa but also underscores the complexity and specificity of microbial interactions within the human gut, particularly in the Chinese population.
    OBJECTIVE: This study presents a system-level map of the differences between feces and rectal mucosal microbial communities in samples with colorectal cancer risk. It reveals the unique microecological characteristics of rectal mucosa and its potential influence on health. Additionally, it provides novel insights into the role of the gut microbiome in the pathogenesis of colorectal cancer and paves the way for the development of new prevention and treatment strategies.
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  • 文章类型: Journal Article
    目的:本研究旨在利用宏基因组测序数据建立肺炎克雷伯菌基因型耐药性检测方法。
    方法:我们利用组装基因组的Lasso回归来鉴定六种抗生素的遗传抗性决定因素(庆大霉素,妥布霉素,亚胺培南,美罗培南,头孢他啶,甲氧苄啶/磺胺甲恶唑)。遗传特征被加权,分组为集群以建立分类器模型。通过整合“可能物种”的新策略确定检测到的抗生素抗性基因(ARG)的起源物种,“基因拷贝数计算”和“物种特异性kmers”。在回顾性案例研究中评估了该方法的性能。
    结果:我们的研究对3928株肺炎克雷伯菌进行了机器学习,对于各种抗生素,产生AUC>0.9的稳定模型。GenseqAMR,基于读取的软件,短读数数据集表现出高精度(AUC0.926-0.956)。物种特异性kmer策略的整合显着提高了ARG物种归因,平均准确率为96.67%。在191个肺炎克雷伯菌阳性临床标本的回顾性研究中(基因组覆盖率为0.68%-93.39%),GenseqAMR平均预测了84.23%的AST结果。它显示了88.76%-96.26%的电阻预测精度,提供基因型AST结果的周转时间(平均±SD:18.34±0.87小时)比基于传统培养的AST(60.15±21.58小时)短。此外,一项涉及63例的回顾性临床病例研究表明,GenseqAMR可导致24例(38.10%)患者的临床治疗发生变化,95.83%(23/24)的这些变化被认为是有益的。
    结论:结论:GenseqAMR是一种用于快速准确预测肺炎克雷伯菌AMR的有前途的工具,通过及时调整抗生素治疗,有可能改善患者的预后。
    OBJECTIVE: The study aimed to develop a genotypic antimicrobial resistance testing method for Klebsiella pneumoniae using metagenomic sequencing data.
    METHODS: We utilized Lasso regression on assembled genomes to identify genetic resistance determinants for six antibiotics (Gentamicin, Tobramycin, Imipenem, Meropenem, Ceftazidime, Trimethoprim/Sulfamethoxazole). The genetic features were weighted, grouped into clusters to establish classifier models. Origin species of detected antibiotic resistant gene (ARG) was determined by novel strategy integrating \"possible species,\" \"gene copy number calculation\" and \"species-specific kmers.\" The performance of the method was evaluated on retrospective case studies.
    RESULTS: Our study employed machine learning on 3928 K. pneumoniae isolates, yielding stable models with AUCs > 0.9 for various antibiotics. GenseqAMR, a read-based software, exhibited high accuracy (AUC 0.926-0.956) for short-read datasets. The integration of a species-specific kmer strategy significantly improved ARG-species attribution to an average accuracy of 96.67%. In a retrospective study of 191 K. pneumoniae-positive clinical specimens (0.68-93.39% genome coverage), GenseqAMR predicted 84.23% of AST results on average. It demonstrated 88.76-96.26% accuracy for resistance prediction, offering genotypic AST results with a shorter turnaround time (mean ± SD: 18.34 ± 0.87 hours) than traditional culture-based AST (60.15 ± 21.58 hours). Furthermore, a retrospective clinical case study involving 63 cases showed that GenseqAMR could lead to changes in clinical treatment for 24 (38.10%) cases, with 95.83% (23/24) of these changes deemed beneficial.
    CONCLUSIONS: In conclusion, GenseqAMR is a promising tool for quick and accurate AMR prediction in Klebsiella pneumoniae, with the potential to improve patient outcomes through timely adjustments in antibiotic treatment.
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  • 文章类型: Letter
    暂无摘要。
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  • 文章类型: Case Reports
    诺卡氏菌病表现出时间分类,包括急性,亚急性,和慢性阶段以及不同的典型定位,如肺部,皮肤,和传播形式。播散性诺卡尼病,通常由诺卡氏菌小行星引起,巴西奈斯,和N.Farcinica,继续导致大量的发病率和死亡率。在这里,我们报道了一例微小病变患者的耳道诺卡氏菌引起的危及生命的播散性诺卡氏菌病.这项研究强调了在临床环境中诊断和治疗未知感染的困难,并强调了实验室在解决由罕见病原体引起的传染病方面发挥的重要作用。
    Nocardiosis demonstrates a temporal categorization that includes acute, subacute, and chronic stages alongside distinct typical localizations such as pulmonary, cutaneous, and disseminated forms. Disseminated nocardiosis, commonly caused by Nocardia asteroides, N. brasiliensis, and N. farcinica, continues to result in substantial morbidity and mortality. Herein, we report a life-threatening disseminated nocardiosis caused by Nocardia otitidiscaviarum in a patient with minimal change disease. This study emphasizes the difficulty in the diagnosis and treatment of unknown infections in clinical settings and highlights the important role played by laboratories in solving infectious diseases caused by rare pathogens.
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  • 文章类型: Journal Article
    抗菌素耐药性(AMR)是全球公共卫生的主要威胁,抗生素抗性基因(ARGs)在人类中广泛分布,动物,和环境。农业环境正在成为ARGs和抗生素抗性细菌(ARB)的关键研究领域。虽然皮肤是ARGs和ARB的重要储库,农业环境和人类皮肤之间的传播机制仍不清楚。先前的研究证实,猪场环境暴露会改变皮肤微生物组,但是这些变化的时间表并不明确。为了提高对这些变化的理解并确定具体时间,我们设计了一项针对猪场工人和学生的队列研究,通过收集皮肤和环境样本,探讨猪场每日职业暴露对人体皮肤微生物组的影响。结果表明,暴露于与牲畜相关的环境中,微生物比学校环境更丰富,可以重塑人类皮肤微生物组和抗生素抗性组。暴露5小时足以通过富集微生物和ARG来改变工人皮肤中的微生物组和ARG结构。这些变化一旦形成就被保留下来。进一步的分析表明,宿主微生物携带的ARG可能在环境与工人皮肤之间转移,并有可能使用农场工人作为ARG载体扩展到普通人群。这些结果引起了人们对ARGs向更广泛社区的潜在传播的担忧。因此,有必要在生产过程中采取相应的干预措施,以减少ARGs和ARB传播的可能性。
    Antimicrobial resistance (AMR) is a major threat to global public health, and antibiotic resistance genes (ARGs) are widely distributed across humans, animals, and environment. Farming environments are emerging as a key research area for ARGs and antibiotic resistant bacteria (ARB). While the skin is an important reservoir of ARGs and ARB, transmission mechanisms between farming environments and human skin remain unclear. Previous studies confirmed that swine farm environmental exposures alter skin microbiome, but the timeline of these changes is ill defined. To improve understanding of these changes and to determine the specific time, we designed a cohort study of swine farm workers and students through collected skin and environmental samples to explore the impact of daily occupational exposure in swine farm on human skin microbiome. Results indicated that exposure to livestock-associated environments where microorganisms are richer than school environment can reshape the human skin microbiome and antibiotic resistome. Exposure of 5 h was sufficient to modify the microbiome and ARG structure in workers\' skin by enriching microorganisms and ARGs. These changes were preserved once formed. Further analysis indicated that ARGs carried by host microorganisms may transfer between the environment with workers\' skin and have the potential to expand to the general population using farm workers as an ARG vector. These results raised concerns about potential transmission of ARGs to the broader community. Therefore, it is necessary to take corresponding intervention measures in the production process to reduce the possibility of ARGs and ARB transmission.
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  • 文章类型: Journal Article
    肠道菌群失调增加了艰难梭菌感染(CDI)的易感性。在这项研究中,我们监测艰难梭菌定植(CDC)患者从无CDC状态(CDN)到CDC状态(CDCp),CDI患者从CDI前无症状状态(PRECDI),CDI状态(ONCDI),CDI后无症状状态(后CDI)。基于宏基因组测序,我们旨在研究肠道菌群与艰难梭菌之间的相互作用模式。CDN和CDCp之间的微生物群多样性没有显着差异。在CDCP中,拟杆菌和产生短链脂肪酸(SCFA)的细菌增加,SCFA产生菌与艰难梭菌定植呈正相关。与PRECDI相比,ONCDI和POSTCDI显示微生物群多样性显著下降,特别是在拟杆菌和产生SCFA的细菌中,机会致病菌与艰难梭菌呈正相关。脂肪酸代谢,氨基酸生物合成富集在CDN中,CDCP,PRECDI,而胆汁分泌在ONCDI和POSTCDI中富集。CDN和CDCp中的微生物群和代谢途径相互作用网络更为复杂,特别是脂肪酸和胆汁酸代谢途径。拟杆菌和产生SCFA的细菌的增加,影响氨基酸和脂肪酸代谢,与对艰难梭菌的定植抗性和抑制CDI的发展有关。
    Gut microbiota dysbiosis increases the susceptibility to Clostridioides difficile infection (CDI). In this study, we monitored C. difficile colonization (CDC) patients from no CDC status (CDN) to CDC status (CDCp) and CDI patients from asymptomatic status before CDI (PRECDI), CDI status (ONCDI), to asymptomatic status after CDI (POSTCDI). Based on metagenomic sequencing, we aimed to investigate the interaction pattern between gut microbiota and C. difficile. There was no significant difference of microbiota diversity between CDN and CDCp. In CDCp, Bacteroidetes and short-chain fatty acid (SCFA)-producing bacteria increased, with a positive correlation between SCFA-producing bacteria and C. difficile colonization. Compared with PRECDI, ONCDI and POSTCDI showed a significant decrease in microbiota diversity, particularly in Bacteroidetes and SCFA-producing bacteria, with a positive correlation between opportunistic pathogen and C. difficile. Fatty acid metabolism, and amino acid biosynthesis were enriched in CDN, CDCp, and PRECDI, while bile secretion was enriched in ONCDI and POSTCDI. Microbiota and metabolic pathways interaction networks in CDN and CDCp were more complex, particularly pathways in fatty acid and bile acid metabolism. Increasing of Bacteroidetes and SCFA-producing bacteria, affecting amino acid and fatty acid metabolism, is associated with colonization resistance to C. difficile and inhibiting the development of CDI.
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  • 文章类型: Journal Article
    背景:微生物群可能与食管鳞状细胞癌(ESCC)的发展有关。然而,目前尚不清楚微生物生物标志物结合流行病学因素对早期发现ESCC和癌前病变的预测价值.
    方法:从中国不同食管状态的349名参与者中收集了449份标本(食管拭子和唾液),通过16SrRNA测序从基因水平到物种水平探索和验证ESCC相关的微生物生物标志物。宏基因组测序和实时定量聚合酶链反应。
    结果:包括放线菌(A.g_1,A.g_2,A.g_3,A.g_4),具核梭杆菌(F.n_1,F.n_2,F.n_3),溶血嗜血杆菌(H.h_1),牙龈卟啉单胞菌(P.g_1、P.g_2、P.g_3)和南方链球菌(S.a_1)通过宏基因组测序进行探索,以早期检测需要组(低级别上皮内瘤变,高级别上皮内瘤变和ESCC)与无这些病变的参与者作为Noneed组。每个微生物靶标都存在显着的定量差异,其中唾液中的检测效率高于食管拭子。在唾液中,基于微生物生物标志物的曲线下面积(AUC)(A.g_4281P.g_3281H.h_1281S.a_1281F.在探索队列中,n_2)为0.722(95%CI0.621-0.823)。结合流行病学因素(年龄,吸烟,饮酒,摄入高温食物和牙痛),在探索队列中,AUC提高到0.869(95%CI0.802-0.937),在验证队列中,AUC为0.757(95%CI0.663-0.852)。
    结论:在中国,将唾液中的微生物生物标志物与流行病学因素相结合,以早期发现ESCC和癌前病变是可行的。
    BACKGROUND: Microbiota may be associated with esophageal squamous cell carcinoma (ESCC) development. However, it is not known the predictive value of microbial biomarkers combining epidemiological factors for the early detection of ESCC and precancerous lesions.
    METHODS: A total of 449 specimens (esophageal swabs and saliva) were collected from 349 participants with different esophageal statuses in China to explore and validate ESCC-associated microbial biomarkers from genes level to species level by 16S rRNA sequencing, metagenomic sequencing and real-time quantitative polymerase chain reaction.
    RESULTS: A bacterial biomarker panel including Actinomyces graevenitzii (A.g_1, A.g_2, A.g_3, A.g_4), Fusobacteria nucleatum (F.n_1, F.n_2, F.n_3), Haemophilus haemolyticus (H.h_1), Porphyromonas gingivalis (P.g_1, P.g_2, P.g_3) and Streptococcus australis (S.a_1) was explored by metagenomic sequencing to early detect the participants in Need group (low-grade intraepithelial neoplasia, high-grade intraepithelial neoplasia and ESCC) vs participants without these lesions as the Noneed group. Significant quantitative differences existed for each microbial target in which the detection efficiency rate was higher in saliva than esophageal swab. In saliva, the area under the curve (AUC) based on the microbial biomarkers (A.g_4 ∩ P.g_3 ∩ H.h_1 ∩ S.a_1 ∩ F.n_2) was 0.722 (95% CI 0.621-0.823) in the exploration cohort. Combining epidemiological factors (age, smoking, drinking, intake of high-temperature food and toothache), the AUC improved to 0.869 (95% CI 0.802-0.937) in the exploration cohort, which was validated with AUC of 0.757 (95% CI 0.663-0.852) in the validation cohort.
    CONCLUSIONS: It is feasible to combine microbial biomarkers in saliva and epidemiological factors to early detect ESCC and precancerous lesions in China.
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