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Ultrasmall epibiont Nanosynbacter lyticus strain TM7x and host bacteria transcriptional activity after initial host parasitism 超小附生纳米溶菌TM7x与寄主初始寄生后寄主细菌转录活性的关系
IF 4.5 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-11-30 DOI: 10.1080/20002297.2023.2287349
Erik L Hendrickson, Batbileg Bor, Kristopher A. Kerns, Lujia Cen, Wenyuan Shi, Xuesong He, Jeffrey S McLean
Oral Saccharibacteria Nanosynbacter lyticus strain TM7× lives as an ultrasmall epibiont on the surface of its host, Schaalia odontolytica strain XH001. Establishing this interaction is a poorly und...
口腔溶牙纳米合杆菌tm7x以超小型表观生物的形式存在于宿主溶牙沙利亚菌XH001的表面。建立这种互动是一个很糟糕的……
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引用次数: 0
Sub-lethal concentrations of chlorhexidine inhibit Candida albicans growth by disrupting ROS and metal ion homeostasis 亚致死浓度的氯己定通过破坏活性氧和金属离子稳态来抑制白色念珠菌的生长
2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-11-09 DOI: 10.1080/20002297.2023.2278937
Qian Jiang, Yuchen Deng, Shuaihu Li, Deqin Yang, Li Tao
Candida albicans is a normal resident of the human oral cavity. It is also the most common fungal pathogen, causing various oral diseases, particularly in immunocompromised individuals. Chlorhexidine digluconate (CHG) is a broad-spectrum antimicrobial agent widely used in dental practice and has been recommended to treat oral candidiasis. However, its action mechanism against the fungal pathogen C. albicans remains poorly understood. The aim of the present study was to investigate the effect of CHG at sub-lethal concentrations against C. albicans. CHG inhibited the growth of C. albicans in a dose- and time-dependent manner. Cells treated with CHG exhibited altered membrane permeability, reduced metabolic activity, and enhanced metal ion and reactive oxygen species (ROS) accumulation. Copper-sensing transcription factor Mac1, iron-sensing transcription factors Sfu1 and Sef2, and copper transporter Ctr1 regulated intracellular metal ion and ROS homeostasis in response to CHG. Deletion of MAC1, SFU1, or SEF2 increased intracellular ROS production and cell susceptibility to CHG. This study revealed a novel mechanism by which CHG induced apoptosis of C. albicans cells through the disruption of metal ion and ROS homeostasis, which may help to identify new targets for fungal infections.
白色念珠菌是人类口腔的正常居民。它也是最常见的真菌病原体,引起各种口腔疾病,特别是在免疫功能低下的个体中。二光酸氯己定(CHG)是一种广泛应用于牙科实践的广谱抗菌药物,已被推荐用于治疗口腔念珠菌病。然而,其对真菌病原体白色念珠菌的作用机制尚不清楚。本研究的目的是探讨亚致死浓度的CHG对白色念珠菌的作用。CHG抑制白色念珠菌生长呈剂量和时间依赖性。经CHG处理的细胞表现出膜通透性改变,代谢活性降低,金属离子和活性氧(ROS)积累增强。铜敏感转录因子Mac1、铁敏感转录因子Sfu1和Sef2以及铜转运体Ctr1调节细胞内金属离子和ROS稳态响应CHG。MAC1、SFU1或SEF2的缺失增加了细胞内ROS的产生和细胞对CHG的易感性。本研究揭示了CHG通过破坏金属离子和ROS稳态诱导白色念珠菌细胞凋亡的新机制,这可能有助于发现真菌感染的新靶点。
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引用次数: 0
Multiomics analysis reveals the genetic and metabolic characteristics associated with the low prevalence of dental caries. 多组学分析揭示了与龋齿低患病率相关的遗传和代谢特征。
IF 3.7 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-11-02 eCollection Date: 2023-01-01 DOI: 10.1080/20002297.2023.2277271
Jinshen Liu, Si-Ying Ye, Xin-Dong Xu, Qiulin Liu, Fei Ma, Xueting Yu, Yu-Hong Luo, Ling-Ling Chen, Xiaojuan Zeng

Background: Despite poor oral hygiene, the Baiku Yao (BKY) ethnic group in China presents a low prevalence of dental caries, which may be related to genetic susceptibility. Due to strict intra-ethnic marriage rule, this ethnic has an advantage in studying the interaction between genetic factors and other regulatory factors related to dental caries.

Methods: Peripheral blood from a caries-free adult male was used for whole genome sequencing, and the BKY assembled genome was compared to the Han Chinese genome. Oral saliva samples were collected from 51 subjects for metabolomic and metagenomic analysis. Multiomics data were integrated for combined analysis using bioinformatics approaches.

Results: Comparative genomic analysis revealed the presence of structural variations in several genes associated with dental caries. Metabolomic and metagenomic sequencing demonstrated the caries-free group had significantly higher concentration of antimicrobials and higher abundance of core oral health-related microbiota. The functional analysis indicated that cationic antimicrobial peptide resistance and the lipopolysaccharide biosynthesis pathway were enriched in the caries-free group.

Conclusions: Our study provided new insights into the specific regulatory mechanisms that contribute to the low prevalence of dental caries in the specific population and may provide new evidence for the genetic diagnosis and control of dental caries.

背景:尽管口腔卫生条件较差,但中国白库瑶族的龋齿患病率较低,这可能与遗传易感性有关。由于严格的种族内部婚姻规则,该种族在研究遗传因素与其他与龋齿相关的调节因素之间的相互作用方面具有优势。方法:采用无龋成年男性外周血进行全基因组测序,并将BKY组装的基因组与汉族基因组进行比较。采集51名受试者的口腔唾液样本进行代谢组学和宏基因组分析。使用生物信息学方法整合多组学数据进行组合分析。结果:比较基因组分析显示,与龋齿相关的几个基因存在结构变异。代谢组学和宏基因组测序表明,无龋组的抗菌药物浓度显著较高,口腔健康相关核心微生物群丰度较高。功能分析表明,阳离子抗菌肽耐药性和脂多糖生物合成途径在无龋组中富集。结论:我们的研究为特定人群中导致龋齿低患病率的特定调控机制提供了新的见解,并可能为龋齿的基因诊断和控制提供新的证据。
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引用次数: 0
Oral microbiota study of the patients after hospitalisation for COVID-19, considering selected dental indices and antibiotic therapy using the next generation sequencing method (NGS). 新冠肺炎患者住院后的口腔微生物群研究,考虑选择的牙科指标和使用下一代测序方法(NGS)的抗生素治疗。
IF 4.5 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-10-11 eCollection Date: 2023-01-01 DOI: 10.1080/20002297.2023.2264591
Barbara Brzychczy-Sroka, Katarzyna Talaga-Ćwiertnia, Agnieszka Sroka-Oleksiak, Artur Gurgul, Elżbieta Zarzecka-Francica, Wojciech Ostrowski, Janusz Kąkol, Joanna Zarzecka, Monika Brzychczy-Włoch

Background: Poor oral hygiene and the increased incidence and severity of periodontitis may exacerbate SARS-CoV-2 infection. The aim was to evaluate the oral microbiota of 60 participants divided into groups: COVID-19 convalescents who received antibiotics during hospitalization (I), COVID-19 convalescents without antibiotic therapy (II) and healthy individuals (III).

Materials and methods: Dental examination was conducted, and oral health status was evaluated using selected dental indexes. Clinical samples (saliva, dorsal swabs, supragingival and subgingival plaque) were collected and used for metagenomic library to the next-generation sequencing (NGS) preparation.

Results: Each of the clinical materials in particular groups of patients showed a statistically significant and quantitatively different bacterial composition. Patients from group I showed significantly worse oral health, reflected by higher average values of dental indexes and also a higher percentage of Veillonella, Tannerella, Capnocytophaga and Selenomonas genera in comparison to other groups. Additionally, a statistically significant decrease in the amount of Akkermansia type in both groups with COVID-19 was observed for all materials.

Conclusions: The primary factor affecting the composition of oral microbiota was not the SARS-CoV-2 infection itself, but the use of antibiotic therapy. The increased percentage of pro-inflammatory pathogens observed in COVID-19 patients underscores the importance of preventing periodontal disease and improving oral hygiene in the future.

背景:口腔卫生不良以及牙周炎发病率和严重程度的增加可能会加剧严重急性呼吸系统综合征冠状病毒2型的感染。目的是评估60名参与者的口腔微生物群,这些参与者分为两组:住院期间服用抗生素的新冠肺炎康复者(I)、未接受抗生素治疗的新冠肺炎康复者(II)和健康个体(III)。材料和方法:进行牙科检查,并使用选定的牙科指标评估口腔健康状况。收集临床样本(唾液、背拭子、龈上和龈下菌斑),用于下一代测序(NGS)制备的宏基因组文库。结果:在特定的患者组中,每种临床材料都显示出具有统计学意义和数量上不同的细菌组成。与其他组相比,第I组患者的口腔健康状况明显较差,表现为牙齿指数的平均值较高,并且韦氏菌属、单核菌属、Capnocytophaga属和Selenomonas属的比例也较高。此外,在所有材料中,观察到新冠肺炎两组患者的Akkermansia类型数量在统计学上显著减少。结论:影响口腔微生物群组成的主要因素不是严重急性呼吸系统综合征冠状病毒2型感染本身,而是抗生素治疗的使用。在新冠肺炎患者中观察到的促炎病原体百分比增加,突显了未来预防牙周病和改善口腔卫生的重要性。
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引用次数: 0
Oral microbiome and ischemic stroke risk among elderly Chinese women. 中国老年妇女的口腔微生物组与缺血性中风风险。
IF 4.5 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-10-08 eCollection Date: 2023-01-01 DOI: 10.1080/20002297.2023.2266655
Cong Wang, Yaohua Yang, Qiuyin Cai, Yutang Gao, Hui Cai, Jie Wu, Wei Zheng, Jirong Long, Xiao-Ou Shu

Background: Stroke, a leading cause of disability worldwide, has been associated with periodontitis. However, whether stroke risk is related to oral microbiota remains unknown. This study aims to evaluate the associations between the oral microbiome and ischemic stroke risk.

Methods: In a case-control study of 134 case-control pairs nested within a prospective cohort study, we examined pre-diagnostic oral microbiome in association with stroke risk via shotgun metagenomic sequencing. The microbial sub-community and functional profiling were performed using Latent Dirichlet Allocation and HUMAnN2. Associations of microbial diversity, sub-community structure, and individual microbial features with ischemic stroke risk were evaluated via conditional logistic regression.

Results: Alpha and beta diversities differ significantly between cases and controls. One genus- and two species-level sub-communities were significantly associated with decreased ischemic stroke risk, with odds ratios (95% confidence intervals) of 0.52 (0.31-0.90), 0.51 (0.31-0.84), and 0.60 (0.36-0.99), respectively. These associations were potentially driven by the representative taxa in these sub-communities, i.e., genus Corynebacterium and Lautropia, and species Lautropia mirabilis and Neisseria elongate (p < 0.05). Additionally, 55 taxa, 1,237 gene families, and 90 metabolic pathways were associated with ischemic stroke risk at p < 0.05.

Conclusion: Our study highlights the role of oral microbiota in the etiology of ischemic stroke and calls for further research.

背景:中风是世界范围内致残的主要原因,与牙周炎有关。然而,中风风险是否与口腔微生物群有关仍然未知。本研究旨在评估口腔微生物组与缺血性中风风险之间的关系。方法:在一项前瞻性队列研究中,我们对134对病例对照配对进行了病例对照研究,通过霰弹枪宏基因组测序检查了与中风风险相关的诊断前口腔微生物组。使用潜在狄利克雷分配和HUMAnN2进行微生物亚群落和功能分析。通过条件logistic回归评估微生物多样性、亚群落结构和个体微生物特征与缺血性卒中风险的相关性。结果:病例和对照组之间的α和β多样性存在显著差异。一属和两种水平的亚群落与缺血性中风风险的降低显著相关,优势比(95%置信区间)分别为0.52(0.31-0.90)、0.51(0.31-0.84)和0.60(0.36-0.99)。这些关联可能是由这些亚群落中的代表性分类群驱动的,即棒状杆菌属和Lautropia,以及奇异LautropiaMirabilis和Neisseria细长种(p p 结论:我们的研究强调了口腔微生物群在缺血性脑卒中病因中的作用,需要进一步研究。
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引用次数: 0
Alterations of oral microbiota are associated with the development and severity of acute pancreatitis. 口腔微生物群的改变与急性胰腺炎的发展和严重程度有关。
IF 4.5 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-10-05 eCollection Date: 2023-01-01 DOI: 10.1080/20002297.2023.2264619
Yiting Liu, Hang Liu, Yuping Rong, Qiao Shi, Qiang Yang, Hanjun Li, Zhengle Zhang, Jing Tao

Acute pancreatitis (AP) is a common abdomen clinical emergency. Most APs have mild clinical symptoms and a good prognosis. However, about 20% of patients develop severe acute pancreatitis (SAP), increasing morbidity and mortality. The microbiome's impact on AP pathophysiology has received increasing attention. Hence, to explore changes in oral microbial composition in acute pancreatitis, we collected clinical information and oral saliva samples from 136 adult participants: 47 healthy controls, 43 acute mild AP (MAP), 29 moderate AP (MSAP), and 17 severe AP (SAP). Using 16S rRNA gene sequencing, 663,175 high-quality sequences were identified. The relative abundance and diversity of oral microorganisms in AP patients increased, with decreased beneficial bacteria such as Streptococcus, Neisseria, and Gemella, and increased Prevotella, Veillonella, Granulicatella, Actinomyces, and Peptostreptococcus in the AP group. Further changes in microbial composition occurred with increasing disease severity, including a decreased abundance of beneficial bacteria such as Neisseria, Haemophilus, and Gemella in MSAP and SAP compared to MAP. Moreover, the Lefse analysis showed that Prevotella, Peptostreptococcus, Actinomyces, and Porphyromonas were better microbial markers for AP. Therefore, oral microbiome changes could distinguish AP from healthy individuals and serve as an early novel predictor of disease severity in AP patients.

急性胰腺炎(AP)是一种常见的腹部临床急症。大多数AP具有轻微的临床症状和良好的预后。然而,大约20%的患者发展为严重急性胰腺炎(SAP),增加了发病率和死亡率。微生物组对AP病理生理学的影响越来越受到关注。因此,为了探索急性胰腺炎口腔微生物组成的变化,我们收集了136名成年参与者的临床信息和口腔唾液样本:47名健康对照、43名急性轻度AP(MAP)、29名中度AP(MSAP)和17名重度AP(SAP)。利用16S rRNA基因测序,鉴定出663175个高质量序列。AP患者口腔微生物的相对丰度和多样性增加,有益菌如链球菌、奈瑟菌和Gemela减少,AP组的普雷沃氏菌、韦氏菌、Granulicatella、放线菌和链球菌增加。微生物组成的进一步变化随着疾病严重程度的增加而发生,包括与MAP相比,MSAP和SAP中有益细菌(如奈瑟菌、嗜血杆菌和Gemella)的丰度降低。此外,Lefse分析表明,普雷沃氏菌、链球菌、放线菌和卟啉单胞菌是AP的较好微生物标记。因此,口腔微生物组的变化可以将AP与健康个体区分开来,并作为AP患者疾病严重程度的早期新预测指标。
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引用次数: 1
Smoking, tooth loss and oral hygiene practices have significant and site-specific impacts on the microbiome of oral mucosal surfaces: a cross-sectional study. 一项横断面研究显示,吸烟、牙齿脱落和口腔卫生习惯对口腔粘膜表面的微生物组有显著的、特定地点的影响。
IF 4.5 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-10-02 eCollection Date: 2023-01-01 DOI: 10.1080/20002297.2023.2263971
Sheila Galvin, Sviatlana Anishchuk, Claire M Healy, Gary P Moran

We investigated bacterial colonisation patterns of healthy mucosa (buccal, tongue, palate and floor of mouth) in a cohort of adults in order to determine how smoking, tooth loss, plaque levels and oral hygiene practices impacted on mucosal colonisation. A total of 322 swabs were recovered from 256 participants, of whom 46% were current smokers. We analysed colonization by sequencing the V1-V3 regions of the 16S rRNA gene. Palate and tongue microbiomes generally exhibited greater biodiversity than buccal and floor of mouth. Although Neisseria, Lautropia and Haemophilus spp. showed reduced abundance in smokers, buccal mucosa specifically showed a significant increase in Prevotella spp., whereas tongue and floor of mouth tended towards increased abundance of Streptococcus spp. Unexpectedly, tooth brushing frequency had a greater impact on mucosal community structure than plaque levels. Tooth loss was associated with significant reductions in mucosal biodiversity and had site-specific impacts, with buccal communities showing increased abundance of periodontitis-associated species and Rothia mucilaginosa, whereas tongue communities exhibited increased abundance of several streptococcal OTUs and reduced abundance of Haemophilus spp. This study highlights the complex relationship between mucosal colonisation and host factors, highlighting the need for careful consideration of these factors in mucosal microbiome studies.

我们调查了一组成年人健康粘膜(颊、舌、腭和口腔底部)的细菌定植模式,以确定吸烟、牙齿脱落、牙菌斑水平和口腔卫生习惯如何影响粘膜定植。共从256名参与者身上采集到322份拭子,其中46%是目前的吸烟者。我们通过对16S rRNA基因的V1-V3区域进行测序来分析定植。腭和舌头的微生物群通常比颊和口腔底部表现出更大的生物多样性。尽管Neisseria、Lautropia和Haemophilus spp.在吸烟者中的丰度降低,但口腔粘膜中普雷沃氏菌的丰度特别显著增加,而舌头和口腔底部的链球菌丰度往往增加。出乎意料的是,刷牙频率对粘膜群落结构的影响比牙菌斑水平更大。牙齿脱落与粘膜生物多样性的显著减少有关,并具有特定地点的影响,口腔群落显示牙周炎相关物种和粘液Rothia mucilaginosa的丰度增加,而舌头群落显示几种链球菌OTU的丰度增加和嗜血杆菌的丰度降低。这项研究强调了粘膜定植与宿主因素之间的复杂关系,强调了在粘膜微生物组研究中仔细考虑这些因素的必要性。
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引用次数: 1
Site-specialization of human oral Gemella species. 人类口腔 Gemella 菌种的部位特化。
IF 3.7 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-06-22 eCollection Date: 2023-01-01 DOI: 10.1080/20002297.2023.2225261
Julian Torres-Morales, Jessica L Mark Welch, Floyd E Dewhirst, Gary G Borisy

Gemella species are core members of the human oral microbiome in healthy subjects and are regarded as commensals, although they can cause opportunistic infections. Our objective was to evaluate the site-specialization of Gemella species among various habitats within the mouth by combining pangenomics and metagenomics. With pangenomics, we identified genome relationships and categorized genes as core and accessory to each species. With metagenomics, we identified the primary oral habitat of individual genomes. Our results establish that the genomes of three species, G. haemolysans, G. sanguinis and G. morbillorum, are abundant and prevalent in human mouths at different oral sites: G. haemolysans on buccal mucosa and keratinized gingiva; G. sanguinis on tongue dorsum, throat, and tonsils; and G. morbillorum in dental plaque. The gene-level basis of site-specificity was investigated by identifying genes that were core to Gemella genomes at a specific oral site but absent from other Gemella genomes. The riboflavin biosynthesis pathway was present in G. haemolysans genomes associated with buccal mucosa but absent from the rest of the genomes. Overall, metapangenomics show that Gemella species have clear ecological preferences in the oral cavity of healthy humans and provides an approach to identifying gene-level drivers of site specificity.

Gemella 菌是健康人口腔微生物组的核心成员,被认为是共生菌,但它们也会引起机会性感染。我们的目的是通过结合泛基因组学和元基因组学,评估 Gemella 菌种在口腔内不同栖息地的专一性。通过庞基因组学,我们确定了基因组关系,并将基因分为核心基因和附属基因。通过元基因组学,我们确定了各个基因组的主要口腔栖息地。我们的研究结果表明,G. haemolysans、G. sanguinis 和 G. morbillorum 这三个物种的基因组在人类口腔的不同口腔部位都很丰富和普遍:G. haemolysans 在颊粘膜和角化的牙龈上;G. sanguinis 在舌背、咽喉和扁桃体上;G. morbillorum 在牙菌斑中。通过识别在特定口腔部位的革兰氏菌基因组中为核心但在其他革兰氏菌基因组中不存在的基因,研究了部位特异性的基因水平基础。核黄素生物合成途径存在于与颊粘膜相关的血凝酵母菌基因组中,但在其他基因组中却不存在。总之,元基因组学表明,Gemella 菌种在健康人的口腔中有明显的生态偏好,并提供了一种方法来识别基因水平上的位点特异性驱动因素。
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引用次数: 0
Fusobacterium nucleatum dissemination by neutrophils. 核分枝杆菌通过中性粒细胞传播。
IF 3.7 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-06-04 eCollection Date: 2023-01-01 DOI: 10.1080/20002297.2023.2217067
Felix Ellett, Nazli I Kacamak, Carla R Alvarez, Eduardo H S Oliveira, Hatice Hasturk, Bruce J Paster, Alpdogan Kantarci, Daniel Irimia

Recent studies uncovered that Fusobacterium nucleatum (Fn), a common, opportunistic bacterium in the oral cavity, is associated with a growing number of systemic diseases, ranging from colon cancer to Alzheimer's disease. However, the pathological mechanisms responsible for this association are still poorly understood. Here, we leverage recent technological advances to study the interactions between Fn and neutrophils. We show that Fn survives within human neutrophils after phagocytosis. Using in vitro microfluidic devices, we determine that human neutrophils can protect and transport Fn over large distances. Moreover, we validate these observations in vivo by showing that neutrophils disseminate Fn using a zebrafish model. Our data support the emerging hypothesis that bacterial dissemination by neutrophils is a mechanistic link between oral and systemic diseases. Furthermore, our results may ultimately lead to therapeutic approaches that target specific host-bacteria interactions, including the dissemination process.

最近的研究发现,口腔中常见的机会性细菌--核酸扶桑分枝杆菌(Fn)与越来越多的全身性疾病(从结肠癌到阿尔茨海默病)有关。然而,人们对造成这种关联的病理机制仍然知之甚少。在这里,我们利用最近的技术进步来研究 Fn 与中性粒细胞之间的相互作用。我们发现 Fn 在被吞噬后会存活在人类中性粒细胞内。利用体外微流控装置,我们确定人类中性粒细胞可以保护并远距离运输 Fn。此外,我们还利用斑马鱼模型证明了中性粒细胞能在体内传播 Fn,从而验证了这些观察结果。我们的数据支持新出现的假说,即中性粒细胞传播细菌是口腔疾病和全身疾病之间的机理联系。此外,我们的研究结果最终可能会导致针对特定宿主-细菌相互作用(包括传播过程)的治疗方法。
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引用次数: 0
Rapid specific detection of oral bacteria using Cas13-based SHERLOCK. 利用基于 Cas13 的 SHERLOCK 快速特异性检测口腔细菌。
IF 3.7 2区 医学 Q2 MICROBIOLOGY Pub Date : 2023-05-11 eCollection Date: 2023-01-01 DOI: 10.1080/20002297.2023.2207336
Jett Liu, Camden Carmichael, Hatice Hasturk, Wenyuan Shi, Batbileg Bor

Decades of ongoing research has established that oral microbial communities play a role in oral diseases such as periodontitis and caries. Yet the detection of oral bacteria and the profiling of oral polymicrobial communities currently rely on methods that are costly, slow, and technically complex, such as qPCR or next-generation sequencing. For the widescale screening of oral microorganisms suitable for point-of-care settings, there exists the need for a low-cost, rapid detection technique. Here, we tailored the novel CRISPR-Cas-based assay SHERLOCK for the species-specific detection of oral bacteria. We developed a computational pipeline capable of generating constructs suitable for SHERLOCK and experimentally validated the detection of seven oral bacteria. We achieved detection within the single-molecule range that remained specific in the presence of off-target DNA found within saliva. Further, we adapted the assay for detecting target sequences directly from unprocessed saliva samples. The results of our detection, when tested on 30 healthy human saliva samples, fully aligned with 16S rRNA sequencing. Looking forward, this method of detecting oral bacteria is highly scalable and can be easily optimized for implementation at point-of-care settings.

数十年的研究证实,口腔微生物群落在牙周炎和龋齿等口腔疾病中起着重要作用。然而,口腔细菌的检测和口腔多微生物群落的分析目前依赖于成本高、速度慢、技术复杂的方法,如 qPCR 或新一代测序。为了大规模筛查适用于护理点环境的口腔微生物,需要一种低成本、快速的检测技术。在这里,我们为口腔细菌的物种特异性检测定制了基于 CRISPR-Cas 的新型检测方法 SHERLOCK。我们开发了一个能够生成适用于 SHERLOCK 的构建体的计算管道,并对七种口腔细菌的检测进行了实验验证。我们实现了单分子范围内的检测,并且在唾液中存在脱靶 DNA 的情况下仍具有特异性。此外,我们还调整了检测方法,以便直接从未经处理的唾液样本中检测目标序列。在对 30 份健康人类唾液样本进行检测时,我们的检测结果与 16S rRNA 测序结果完全一致。展望未来,这种检测口腔细菌的方法具有很强的可扩展性,可以很容易地进行优化,以便在护理点环境中实施。
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引用次数: 0
期刊
Journal of Oral Microbiology
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