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Nanopore-based enrichment of antimicrobial resistance genes - a case-based study. 基于纳米孔的抗菌药耐药性基因富集--一项基于案例的研究。
Pub Date : 2023-01-25 eCollection Date: 2023-01-01 DOI: 10.46471/gigabyte.75
Adrian Viehweger, Mike Marquet, Martin Hölzer, Nadine Dietze, Mathias W Pletz, Christian Brandt

Rapid screening of hospital admissions to detect asymptomatic carriers of resistant bacteria can prevent pathogen outbreaks. However, the resulting isolates rarely have their genome sequenced due to cost constraints and long turn-around times to get and process the data, limiting their usefulness to the practitioner. Here we used real-time, on-device target enrichment ("adaptive") sequencing as a highly multiplexed assay covering 1,147 antimicrobial resistance genes. We compared its utility against standard and metagenomic sequencing, focusing on an isolate of Raoultella ornithinolytica harbouring three carbapenemases (NDM, KPC, VIM). Based on this experimental data, we then modelled the influence of several variables on the enrichment results and predicted the large effect of nucleotide identity (higher is better) and read length (shorter is better). Lastly, we showed how all relevant resistance genes are detected using adaptive sequencing on a miniature ("Flongle") flow cell, motivating its use in a clinical setting to monitor similar cases and their surroundings.

对入院患者进行快速筛查,检测出无症状的耐药细菌携带者,可以防止病原体爆发。然而,由于成本限制以及获取和处理数据的周转时间过长,由此产生的分离物很少进行基因组测序,从而限制了它们对从业人员的有用性。在这里,我们使用实时、设备上目标富集("自适应")测序作为一种高度复用的检测方法,涵盖了 1,147 个抗菌药耐药性基因。我们将其用途与标准测序和元基因组测序进行了比较,重点研究了携带三种碳青霉烯酶(NDM、KPC、VIM)的鸟疫杆菌分离物。根据这些实验数据,我们模拟了几个变量对富集结果的影响,并预测了核苷酸同一性(越高越好)和读取长度(越短越好)的巨大影响。最后,我们展示了如何在微型("Flongle")流式细胞上使用自适应测序技术检测所有相关的耐药基因,从而推动其在临床环境中的应用,以监测类似病例及其周围环境。
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引用次数: 0
Improvements to the Gulf pipefish Syngnathus scovelli genome 海湾管鱼(Syngnathus scovelli)基因组的改良
Pub Date : 2023-01-20 DOI: 10.1101/2023.01.23.525209
B. Ramesh, CM Small, H. Healey, B. Johnson, E. Barker, M. Currey, S. Bassham, M. Myers, WA Cresko, Ag Jones
The Gulf pipefish Syngnathus scovelli has emerged as an important species in the study of sexual selection, development, and physiology, among other topics. The fish family Syngnathidae, which includes pipefishes, seahorses, and seadragons, has become an increasingly attractive target for comparative research in ecological and evolutionary genomics. These endeavors depend on having a high-quality genome assembly and annotation. However, the first version of the S. scovelli genome assembly was generated by short-read sequencing and annotated using a small set of RNA-sequence data, resulting in limited contiguity and a relatively poor annotation. Here, we present an improved genome assembly and an enhanced annotation, resulting in a new official gene set for S. scovelli. By using PacBio long-read high-fidelity (Hi-Fi) sequences and a proximity ligation (Hi-C) library, we fill small gaps and join the contigs to obtain 22 chromosome-level scaffolds. Compared to the previously published genome, the gaps in our novel genome assembly are smaller, the N75 is much larger (13.3 Mb), and this new genome is around 95% BUSCO complete. The precision of the gene models in the NCBI’s eukaryotic annotation pipeline was enhanced by using a large body of RNA-Seq reads from different tissue types, leading to the discovery of 28,162 genes, of which 8,061 were non-coding genes. This new genome assembly and the annotation are tagged as a RefSeq genome by NCBI and thus provide substantially enhanced genomic resources for future research involving S. scovelli.
墨西哥湾管鱼Syngnathus scovelli已成为性别选择、发育和生理学等研究领域的重要物种。包括管鱼、海马和海龙在内的鱼类科Syngnathidae已成为生态和进化基因组学比较研究的一个越来越有吸引力的目标。这些努力依赖于高质量的基因组组装和注释。然而,第一个版本的S.scovelli基因组组装是通过短读测序生成的,并使用一小组RNA序列数据进行注释,导致邻接性有限,注释相对较差。在这里,我们提出了一个改进的基因组组装和增强的注释,从而为S.scovelli产生了一个新的官方基因集。通过使用PacBio长读高保真(Hi-Fi)序列和邻近连接(Hi-C)文库,我们填补了小间隙并连接了重叠群,获得了22个染色体水平的支架。与之前发表的基因组相比,我们新基因组组装中的缺口更小,N75要大得多(13.3Mb),并且这个新基因组的BUSCO完整率约为95%。通过使用来自不同组织类型的大量RNA-Seq读数,NCBI真核注释管道中基因模型的精度得到了提高,从而发现了28162个基因,其中8061个是非编码基因。这种新的基因组组装和注释被NCBI标记为RefSeq基因组,从而为未来涉及S.scovelli的研究提供了显著增强的基因组资源。
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引用次数: 1
Distribution of mosquitoes (Diptera: Culicidae) in Thailand: a dataset. 泰国蚊类分布(双翅目:库蚊科):a dataset。
Pub Date : 2023-01-01 DOI: 10.46471/gigabyte.90
Chutipong Sukkanon, Wannapa Suwonkerd, Kanutcharee Thanispong, Manop Saeung, Pairpailin Jhaiaun, Suntorn Pimnon, Kanaphot Thongkhao, Sylvie Manguin, Theeraphap Chareonviriyaphap

Mosquitoes play a crucial role as primary vectors for various infectious diseases in Thailand. Therefore, accurate distribution information is vital for effectively combating and better controlling mosquito-borne diseases. Here, we present a curated dataset of the mosquito distribution in Thailand comprising 12,278 records of at least 117 mosquito species (Diptera: Culicidae). The main genera included in the dataset are Aedes, Anopheles, Armigeres, Culex, and Mansonia. From 2007 to 2023, data were collected through routine mosquito surveillance and research projects from 1,725 locations across 66 (out of 77) Thai provinces. The majority of the data were extracted from a Thai database of the Thailand Malaria Elimination Program. To facilitate broader access to mosquito-related data and support further exploration of the Thai mosquito fauna, the data were translated into English. Our dataset has been published in the Global Biodiversity Information Facility, making it available for researchers worldwide.

在泰国,蚊子作为各种传染病的主要媒介发挥着至关重要的作用。因此,准确的分布信息对于有效防治和更好地控制蚊媒疾病至关重要。在这里,我们提出了一个泰国蚊子分布的精心整理的数据集,其中包括至少117种蚊子(双翅目:库蚊科)的12,278条记录。数据集中的主要属有伊蚊、按蚊、阿蚊、库蚊和曼蚊。从2007年至2023年,通过常规蚊虫监测和研究项目从泰国66个省(77个省中的66个省)的1725个地点收集了数据。大多数数据是从泰国疟疾消除计划的泰国数据库中提取的。为了方便更广泛地获取与蚊子有关的数据,并支持对泰国蚊子区系的进一步探索,这些数据被翻译成英文。我们的数据集已经发表在全球生物多样性信息设施上,供全世界的研究人员使用。
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引用次数: 0
HTGQC and shinyHTGQC: an R package and shinyR application for quality controls of HTG EDGE-seq protocols. HTGQC 和 shinyHTGQC:用于 HTG EDGE-seq 协议质量控制的 R 软件包和 shinyR 应用程序。
Pub Date : 2022-12-02 eCollection Date: 2022-01-01 DOI: 10.46471/gigabyte.74
Lodovico Terzi di Bergamo, Francesca Guidetti, Davide Rossi, Francesco Bertoni, Luciano Cascione

Extraction-free HTG EdgeSeq protocols are used to profile sets of genes and measure their expression. Thus, these protocols are frequently used to characterise tumours and their microenvironments. However, although positive and control genes are provided, little indication is given concerning the assessment of the technical success of each sample within the sequencing run. We developed HTGQC, an R package for the quality control of HTG EdgeSeq protocols. Additionally, shinyHTGQC is a shiny application for users without computing knowledge, providing an easy-to-use interface for data quality control and visualisation. Quality checks can be performed on the raw sequencing outputs, and samples are flagged as FAIL or ALERT based on the expression levels of the positive and negative control genes.

Availability & implementation: The code is freely available at https://github.com/LodovicoTerzi/HTGQC (R package) and https://lodovico.shinyapps.io/shinyHTGQC/ (shiny application), including test datasets.

免提取 HTG EdgeSeq 协议用于描述基因集并测量其表达。因此,这些方案常用于描述肿瘤及其微环境的特征。然而,虽然提供了阳性基因和对照基因,但几乎没有说明如何评估测序运行中每个样本的技术成功率。我们开发了 HTGQC,这是一个用于 HTG EdgeSeq 方案质量控制的 R 软件包。此外,shinyHTGQC 是一款面向无计算知识用户的shiny应用程序,为数据质量控制和可视化提供了一个易于使用的界面。可对原始测序输出进行质量检查,并根据阳性和阴性对照基因的表达水平将样本标记为FAIL或ALERT:代码可在 https://github.com/LodovicoTerzi/HTGQC(R 软件包)和 https://lodovico.shinyapps.io/shinyHTGQC/(shiny 应用程序)上免费获取,包括测试数据集。
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引用次数: 0
PhysiPKPD: A pharmacokinetics and pharmacodynamics module for PhysiCell. PhysiPKPD:PhysiCell 的药代动力学和药效学模块。
Pub Date : 2022-11-30 eCollection Date: 2022-01-01 DOI: 10.46471/gigabyte.72
Daniel Bergman, Lauren Marazzi, Mukti Chowkwale, Deepa Maheshvare M, Supriya Bidanta, Tarunendu Mapder, Jialun Li

Pharmacokinetics and pharmacodynamics (PKPD) are key considerations in any study of molecular therapies. It is thus imperative to factor their effects into any in silico model of biological tissue involving such therapies. Furthermore, creating a standardized and flexible framework will benefit the community by increasing access to such modules and enhancing their communicability. PhysiCell is an open-source physics-based cell simulator, i.e., a platform for modeling biological tissue, that is quickly being adopted and utilized by the mathematical biology community. We present here PhysiPKPD, an open-source PhysiCell-based package that allows users to include PKPD in PhysiCell models.

Availability & implementation: The source code for PhysiPKPD is located here: https://github.com/drbergman/PhysiPKPD.

药代动力学和药效动力学(PKPD)是分子疗法研究的关键因素。因此,在任何涉及此类疗法的生物组织硅学模型中都必须考虑到它们的影响。此外,创建一个标准化和灵活的框架将使社区受益,增加对此类模块的访问并增强其可交流性。PhysiCell 是一个开源的基于物理学的细胞模拟器,即生物组织建模平台,正迅速被数学生物学界采用和利用。我们在此介绍 PhysiPKPD,这是一个基于 PhysiCell 的开源软件包,允许用户在 PhysiCell 模型中加入 PKPD:PhysiPKPD 的源代码位于:https://github.com/drbergman/PhysiPKPD。
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引用次数: 0
The first complete mitochondrial genome of Diadema antillarum (Diadematoida, Diadematidae). Diadematoida, Diadematidae)的首个完整线粒体基因组。
Pub Date : 2022-11-22 eCollection Date: 2022-01-01 DOI: 10.46471/gigabyte.73
Audrey J Majeske, Alejandro J Mercado Capote, Aleksey Komissarov, Anna Bogdanova, Nikolaos V Schizas, Stephanie O Castro Márquez, Kenneth Hilkert, Walter Wolfsberger, Tarás K Oleksyk

The mitochondrial genome of the long-spined black sea urchin, Diadema antillarum, was sequenced using Illumina next-generation sequencing technology. The complete mitogenome is 15,708 bp in length, containing two rRNA, 22 tRNA and 13 protein-coding genes, plus a noncoding control region of 133 bp. The nucleotide composition is 18.37% G, 23.79% C, 26.84% A and 30.99% T. The A + T bias is 57.84%. Phylogenetic analysis based on 12 complete mitochondrial genomes of sea urchins, including four species of the family Diadematidae, supported familial monophyly; however, the two Diadema species, D. antillarum and D. setosum were not recovered as sister taxa.

利用 Illumina 下一代测序技术对长棘黑海胆 Diadema antillarum 的线粒体基因组进行了测序。完整的线粒体基因组全长 15,708 bp,包含 2 个 rRNA、22 个 tRNA 和 13 个编码蛋白质的基因,以及一个 133 bp 的非编码控制区。核苷酸组成为 18.37% G、23.79% C、26.84% A 和 30.99% T。基于 12 个完整的海胆线粒体基因组(包括 Diadematidae 科的 4 个物种)的系统进化分析支持家族单系性;但是,两个 Diadema 物种 D. antillarum 和 D. setosum 没有恢复为姊妹类群。
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引用次数: 0
A highly contiguous, scaffold-level nuclear genome assembly for the fever tree (Cinchona pubescens Vahl) as a novel resource for Rubiaceae research. 发热树(Cinchona pubescens Vahl)高度连续的支架级核基因组组装,是茜草科研究的新资源。
Pub Date : 2022-10-06 eCollection Date: 2022-01-01 DOI: 10.46471/gigabyte.71
Nataly Allasi Canales, Oscar A Pérez-Escobar, Robyn F Powell, Mats Töpel, Catherine Kidner, Mark Nesbitt, Carla Maldonado, Christopher J Barnes, Nina Rønsted, Natalia A S Przelomska, Ilia J Leitch, Alexandre Antonelli

The Andean fever tree (Cinchona L.; Rubiaceae) is a source of bioactive quinine alkaloids used to treat malaria. C. pubescens Vahl is a valuable cash crop within its native range in northwestern South America, however, genomic resources are lacking. Here we provide the first highly contiguous and annotated nuclear and plastid genome assemblies using Oxford Nanopore PromethION-derived long-read and Illumina short-read data. Our nuclear genome assembly comprises 603 scaffolds with a total length of 904 Mbp (∼82% of the full genome based on a genome size of 1.1 Gbp/1C). Using a combination of de novo and reference-based transcriptome assemblies we annotated 72,305 coding sequences comprising 83% of the BUSCO gene set and 4.6% fragmented sequences. Using additional plastid and nuclear datasets we place C. pubescens in the Gentianales order. This first genomic resource for C. pubescens opens new research avenues, including the analysis of alkaloid biosynthesis in the fever tree.

安第斯热树(Cinchona L.;茜草科)是生物活性奎宁生物碱的来源,可用于治疗疟疾。C. pubescens Vahl 是其原产地南美洲西北部的一种有价值的经济作物,但缺乏基因组资源。在这里,我们利用牛津纳米孔公司 PromethION 衍生的长读数和 Illumina 的短读数数据,首次提供了高度连续且有注释的核基因组和质粒基因组组装。我们的核基因组组装包括 603 个支架,总长度为 904 Mbp(根据 1.1 Gbp/1C 的基因组大小计算,占全基因组的 82%)。我们结合使用从头装配和基于参考的转录组装配,注释了 72,305 个编码序列,其中包括 83% 的 BUSCO 基因组和 4.6% 的片段序列。利用其他质体和核数据集,我们将 C. pubescens 列入龙胆目。这一首个龙胆属植物基因组资源开辟了新的研究途径,包括分析发热树中生物碱的生物合成。
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引用次数: 0
svaRetro and svaNUMT: modular packages for annotating retrotransposed transcripts and nuclear integration of mitochondrial DNA in genome sequencing data. svaRetro 和 svaNUMT:用于注释基因组测序数据中线粒体 DNA 的逆转录转录本和核整合的模块化软件包。
Pub Date : 2022-10-05 eCollection Date: 2022-01-01 DOI: 10.46471/gigabyte.70
Ruining Dong, Daniel Cameron, Justin Bedo, Anthony T Papenfuss

Nuclear integration of mitochondrial genomes and retrocopied transcript insertion are biologically important but often-overlooked aspects of structural variant (SV) annotation. While tools for their detection exist, these typically rely on reanalysis of primary data using specialised detectors rather than leveraging calls from general purpose structural variant callers. Such reanalysis potentially leads to additional computational expense and does not take advantage of advances in general purpose structural variant calling. Here, we present svaRetro and svaNUMT; R packages that provide functions for annotating novel genomic events, such as nonreference retrocopied transcripts and nuclear integration of mitochondrial DNA. The packages were developed to work within the Bioconductor framework. We evaluate the performance of these packages to detect events using simulations and public benchmarking datasets, and annotate processed transcripts in a public structural variant database. svaRetro and svaNUMT provide modular, SV-caller agnostic tools for downstream annotation of structural variant calls.

线粒体基因组的核整合和逆转录转录本插入是结构变异(SV)注释的重要生物学方面,但往往被忽视。虽然已有用于检测它们的工具,但这些工具通常依赖于使用专用检测器对原始数据进行再分析,而不是利用通用结构变异调用器的调用。这种重新分析可能会导致额外的计算费用,而且无法利用通用结构变异调用的进步。在此,我们介绍了 svaRetro 和 svaNUMT;这两个 R 软件包提供了注释新基因组事件的功能,如非参考反转录本和线粒体 DNA 的核整合。这些软件包是在 Bioconductor 框架内开发的。我们利用模拟和公共基准数据集评估了这些软件包检测事件的性能,并在公共结构变异数据库中注释了处理过的转录本。svaRetro 和 svaNUMT 为结构变异调用的下游注释提供了模块化、与 SV 调用器无关的工具。
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引用次数: 0
A phased, chromosome-scale genome of 'Honeycrisp' apple (Malus domestica). 蜜脆 "苹果(Malus domestica)的分阶段染色体级基因组。
Pub Date : 2022-09-19 eCollection Date: 2022-01-01 DOI: 10.46471/gigabyte.69
Awais Khan, Sarah B Carey, Alicia Serrano, Huiting Zhang, Heidi Hargarten, Haley Hale, Alex Harkess, Loren Honaas

The apple cultivar 'Honeycrisp' has superior fruit quality traits, cold hardiness, and disease resistance, making it a popular breeding parent. However, it suffers from several physiological disorders, production, and postharvest issues. Despite several available apple genome sequences, understanding of the genetic mechanisms underlying cultivar-specific traits remains lacking. Here, we present a highly contiguous, fully phased, chromosome-level genome of 'Honeycrisp' apples, using PacBio HiFi, Omni-C, and Illumina sequencing platforms, with two assembled haplomes of 674 Mbp and 660 Mbp, and contig N50 values of 32.8 Mbp and 31.6 Mbp, respectively. Overall, 47,563 and 48,655 protein-coding genes were annotated from each haplome, capturing 96.8-97.4% complete BUSCOs in the eudicot database. Gene family analysis reveals most 'Honeycrisp' genes are assigned into orthogroups shared with other genomes, with 121 'Honeycrisp'-specific orthogroups. This resource is valuable for understanding the genetic basis of important traits in apples and related Rosaceae species to enhance breeding efforts.

苹果栽培品种 "Honeycrisp "具有优良的果实品质特征、耐寒性和抗病性,因此成为广受欢迎的育种亲本。然而,它也存在一些生理失调、生产和采后问题。尽管已有多个苹果基因组序列,但对栽培品种特有性状的遗传机制仍然缺乏了解。在这里,我们利用 PacBio HiFi、Omni-C 和 Illumina 测序平台,展示了高度连续、全相位、染色体级的'Honeycrisp'苹果基因组,其中两个组装的单体分别为 674 Mbp 和 660 Mbp,连接 N50 值分别为 32.8 Mbp 和 31.6 Mbp。总体而言,每个单倍组分别注释了 47563 和 48655 个蛋白质编码基因,捕获了 96.8%-97.4% 完整的裸子植物数据库中的 BUSCOs。基因家族分析表明,大多数 "Honeycrisp "基因被归入与其他基因组共享的正交组,其中有 121 个 "Honeycrisp "特异正交组。这一资源对于了解苹果和相关蔷薇科植物重要性状的遗传基础,从而加强育种工作非常有价值。
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引用次数: 0
PhysiPKPD: A pharmacokinetics and pharmacodynamics module for PhysiCell PhysiPKPD:PhysicCell的药代动力学和药效学模块
Pub Date : 2022-09-15 DOI: 10.1101/2022.09.12.507681
Daniel R. Bergman, Lauren Marazzi, Mukti Chowkwale, Deepa Maheshvare M, Supriya Bidanta, T. Mapder, Jialun Li
Pharmacokinetics and pharmacodynamics are key considerations in any study of molecular therapies. It is thus imperative to factor their effects in to any in silico model of biological tissue involving such therapies. Furthermore, creation of a standardized and flexible framework will benefit the community by increasing access to such modules and enhancing their communicability. PhysiCell is an open source physics-based cell simulator, i.e. a platform for modeling biological tissue, that is quickly being adopted and utilized by the mathematical biology community. We present here PhysiPKPD, an open source PhysiCell-based package that allows users to include PKPD in PhysiCell models. Availability & Implementation The source code for PhysiPKPD is located here: https://github.com/drbergman/PhysiPKPD.
药代动力学和药效学是任何分子治疗研究的关键考虑因素。因此,必须将其影响因素纳入涉及此类治疗的任何生物组织的计算机模型。此外,创建一个标准化和灵活的框架将通过增加对这些模块的访问并增强其可交流性而使社区受益。PhysiCell是一个开源的基于物理的细胞模拟器,即一个模拟生物组织的平台,它正在被数学生物学社区迅速采用和利用。我们在这里介绍PhysiPKPD,一个基于PhysiCell的开源包,允许用户在PhysiCell模型中包含PKPD。可用性和实现PhysiPKPD的源代码位于这里:https://github.com/drbergman/PhysiPKPD。
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引用次数: 3
期刊
GigaByte (Hong Kong, China)
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