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73 results for “pathogen detection”
Simulated NGS datasets for real-time detection of novel pathogens
<p>Datasets based on the <a href="https://doi.org/10.5281/zenodo.3678563">bacterial</a> and <a href="https://doi.org/10.5281/zenodo.4312525">viral</a> simulated NGS datasets. Fastq files correspond to tests sets of those datasets. Basecall files were generated based on the fastq files with an 8nt simulated barcode between the mates of a read pair. The "rn" datasets containg random length subreads (25-250bp) of the original validation and training reads.</p> <p>The Nanopore datasets were resimulated with <a href="https://github.com/liyu95/DeepSimulator">DeepSimulator 1.5</a> (Li et al., 2020) based on the original datasets (i.e. using the same species composition as the original data). The test Nanopore dataset contains full reads (target average length: 8kb) and the training and validation datasets - 250bp subreads.</p>
Dataset for publication: An inter-laboratory study characterizes the impact of bioinformatic approaches on genome-based cluster detection for foodborne bacterial pathogens
<p>This dataset is part of a dry-lab interlaboratory study conducted across Germany, regarding bacterial outbreak detection based on NGS data, with a focus on bioinformatic analysis of four species to identify potential variability caused by different data analysis approaches and human interpretation. Participants were asked to follow their usual in-house protocols while adhering to the general guidelines. A quality assessment (with sample exclusion) was followed by 7-gene Multilocus-Sequence Typing (MLST), core genome Multilocus Sequencing Typing (cgMLST), and SNP calling. The participants were then asked to identify clusters. The study was not intended to resemble a standard proficiency test with a passing/failing grade, but rather to investigate and quantify obvious variability in the results and, where possible, the reasons for it. For this purpose, the datasets included borderline cases in terms of quality.</p>
Fig. 1 in Detection of Escherichia fergusonii - an emerging pathogen harbouring drug resistant genes from seafood samples of Tamil Nadu, India
Fig. 1 — Gene specific PCR amplification of Escherichia fergusonii (lane 1 – 100 bp DNA ladder, lane 2 – positive control (clinical E. fergusonii), lane 3 – negative control, lane 4 – E011, lane 5 – E060)
Dataset for Multiplex-PCR detection and Nanopore-based genotyping of fish pathogens
<p>This is a revised zip file contains scripts, initial fastq files, assembled amplicon (public and from this study) as well as bioinformatics intermediate files used for this study.</p> <p>Changelog:</p> <p>1. Fixed a bug in the 02_consensus.sh to enable proper removal of amplicons with zero depth</p> <p>2. Added a script (06_unclassified_read.sh) to extract and annotate reads that previously could not align to the 4 reference gene segment. Now the previously unclassified reads will be re-align (raw fastq) back to the gene segments as well as an additional tilapia genome assembly to gauge amount of reads mapping to the host genome. Furthermore, any read that still fail to align with minimap2 was subsequently aligned using blastn (-word_size 15 -evalue 0.01) against the same sequences.</p> <p>File Structure and Descriptions</p> <p>├── 01_process.sh : primer trimming, length-based filtering, read alignment, alignment filtering (unique hit) and extraction of uniquely hit reads for consensus generation<br> ├── 02_consensus.sh : [need artic conda env] Generation of consensus based on uniquely-mapped reads and minimal read depth of 20x required to call a variant (or it will be masked)<br> ├── 03_cleanup.sh: General folder and intermediate file re-organization<br> ├── 04_filter.sh: [need quast conda env] statistic of consensus generated and filtering of consensus with one or more ambiguous base (N), not suitable for haplotype<br> ├── 05_cluster.sh: clustering of consensus based on 100% identity threshold to generate putative haplotype<br> ├── 06_unclassified_read.sh: Extraction and annotation of unclassified reads using lenient criteria and with host reference genome as added reference<br> ├── Amplicon_FastQ folder: uniquely mapped fastq files for consensus generation<br> ├── BAM: alignment files generated from minimap2 used as input for the artic pipeline to identify variants<br> ├── Cluster_Rep.txt: Consensus sequences that were chosen to represent each haplotype<br> ├── Consensus folder: consensus fasta files generated for each sample containing sequences for each specific pathogen<br> ├── Coverage folder: coverage and base-level read depth for each sample and each pathogen reference genes<br> ├── Filter: individual fasta sequences (only 1 sequence per file) for each pathogen and each sample without any ambiguous base for subsequent clustering analysis<br> ├── Full_Haplotype.fasta: all possible haplotype sequences generated for each pathogen<br> ├── Gap_Analysis.tsv: Table with percentage of gap (0-100%) for each consensus sequence generated (used for filtering)<br> ├── Haplotype folder: Intermediate file and sample-level haplotype used to infer final haplotype and generate haplotype summary<br> ├── Haplotype_summary.tsv: Table with sample ID and their respectively pathogen haplotype<br> ├── Minimap2_PAF: Intermediate alignment generated from minimap2 used to generate the count table<br> ├── FailMinimap2 folder: FastQ files that didn't align using minimap2. Will be subsequently aligned using blastN (more sensitive) against the same reference sequences as minimap2<br> ├── Host_4Pathogen.fasta: Fasta file containing the tilapia genome and 4 pathogen (primer binding site included)<br> ├── Original: fastq with original naming prior to renaming based on sampleID. a script (rename.sh) was included to show renaming scheme<br> ├── primer.fasta: Primer sequences used for identifying and trimming reads with flanking primer sequence<br> ├── primer.fasta.fai: the index file for primer.fasta<br> ├── PrimerTrim folder: Primer-trimmed reads<br> ├── quast_results: consensus statistics generated by quast<br> ├── RawCount.tsv: Count table generated that can used as a input to generate figure<br> ├── RawFastq folder: Raw reads that have been renamed to reflect sample information<br> ├── readme.md: The current readme file<br> ├── ref_full_latest.fasta: Reference sequence of (gene segments) 4 pathogens e.g. TilV, ISKNV, SAG (Streptococcus agalactiae), FNO (Francisella noatunensis subsp. orientalis)<br> ├── ref_full_latest.primer.fasta: Same as above but with their primer binding sequence trimmed similar to the processed reads<br> ├── ref_full_latest.primer.fasta.fai<br> ├── RenameHaplotype: Script to perform reorganization of cdhit output<br> ├── Seq.stat.tsv: Sequencing statistics<br> ├── Uniq_PAF: Minimap2 alignment file for raw reads that initially failed quality check (no primer present and/or less than 80% query coverage / not unique alignment)<br> ├── Unmap: Raw reads that initial failed quality check (no primer on both ends / less than 80% query coverage / not unique alignment) <br> └── VCF: VCF files from medaka variant calling used to generate the final consensus</p>
Single-molecule Fluorescent In Situ Hybridization (smFISH) for RNA detection in the fungal pathogen Candida albicans dataset
<p><strong>This dataset is connected to the protocol article titled:</strong></p> <p>Single-molecule Fluorescent <em>In Situ</em> Hybridization (smFISH) for RNA detection in the fungal pathogen <em>Candida albicans</em></p> <p><strong>Abstract:</strong></p> <p><em>Candida albicans</em> is the most prevalent human fungal pathogen. Its pathogenicity is linked to the ability of <em>C. albicans</em> to reversibly change morphology and to grow as yeast, pseudohyphal or hyphal cells in response to environmental stimuli. Understanding the molecular regulation controlling those morphological switches remains a challenge that, if solved, could help fight <em>C. albicans</em> infections.</p> <p>While numerous studies investigated gene expression changes occurring during <em>C. albicans</em> morphological switches using bulk approaches (e.g., RNA sequencing), here we describe a single-cell and single-molecule RNA imaging and analysis protocol to measure absolute mRNA counts in morphologically intact cells. To detect endogenous mRNAs in single fixed cells, we optimized a single molecule fluorescent <em>in situ</em> hybridization (smFISH) protocol for <em>C. albicans</em>, which allows one to quantify the differential expression of mRNAs in yeast, pseudohyphae or hyphal cells. We quantified the expression of two mRNAs, cell cycle-controlled mRNA (<em>CLB2)</em> and a transcription regulator (<em>EFG1</em>), which show differential expression in the different morphological cell types and in different nutrient conditions. In this protocol we described in detail the major steps of this approach: growth and fixation, hybridization, imaging, cell-segmentation and mRNA spot analysis. Raw data is provided with the protocol to favour reproducibility. This approach could benefit the molecular characterization of <em>C. albicans</em> and other filamentous fungi, pathogenic or non-pathogenic.</p> <p><strong>Data description:</strong></p> <p>This dataset consists of a FISH experiment spanning two different mRNAs, EFG1 and CLB2, and two different nutrient condition, being SPIDER37 and TSB37 in Candida albicans. For culturing, the C. albicans wildtype strain SC5314 was inoculated at 30 degrees overnight (~15 hours) in 10 mL of TSB medium in a 30 degree (celsius) shaking incubator. Next, samples were diluted to a density of 10^5 cells/ mL and inoculated for 6 hours in either 30 mL TSB medium or Spider medium at 37 degrees in falcon tubes on an orbital microplate shaker. Then, samples were fixated by adding PFA to a final concentration of 4% to the medium. For hybridization, both mRNAs were hybridized independently by specific DNA oligo labelled with a Quasar670 dye to enable the visualisation of single mRNA molecules. As both genes are labelled by the same dye, these oligos were not co-applied to the same sample but to independent samples.</p> <p><strong>Microscopy</strong></p> <p>For smFISH imaging we use an Olympus BX-63 epifluorescence microscope equipped with Ultrasonic stage and UPlanApo 100x 1.35NA oil-immersion objective (Olympus). Lumencore SOLA FISH light source, a Hamamatsu ORCA-Fusion sCMOS camera (6.5 µm-pixel size) mounted using U-CMT C-Mount Adapter, and zero-pixel shift filter sets: F36-500 DAPI HC Brightline Bandpass Filter, F36-502 FITC HC BrightLine Filter, F36-542 Cy3 HC BrightLine Filter, and F36-523 Cy5 HC BrightLine Filter. Images are acquired across 61-81 optical sections (depending on the sample thickness) with a z-step size of 0.2 μm. The CellSens software (Olympus) is used for instrument control and image acquisition. For the DAPI channel 10-50 ms of exposure was used. Whilst, for the CY5 channel, used imaging the FISH probes, 750 ms was applied. </p> <p> </p>
Single-molecule Fluorescent In Situ Hybridization (smFISH) for RNA detection in the fungal pathogen Candida albicans small example dataset
<p><strong>This small example dataset is connected to the protocol article titled:</strong></p> <p>Single-molecule Fluorescent <em>In Situ</em> Hybridization (smFISH) for RNA detection in the fungal pathogen <em>Candida albicans</em></p> <p><strong>Abstract:</strong></p> <p><em>Candida albicans</em> is the most prevalent human fungal pathogen. Its pathogenicity is linked to the ability of <em>C. albicans</em> to reversibly change morphology and to grow as yeast, pseudohyphal or hyphal cells in response to environmental stimuli. Understanding the molecular regulation controlling those morphological switches remains a challenge that, if solved, could help fight <em>C. albicans</em> infections.</p> <p>While numerous studies investigated gene expression changes occurring during <em>C. albicans</em> morphological switches using bulk approaches (e.g., RNA sequencing), here we describe a single-cell and single-molecule RNA imaging and analysis protocol to measure absolute mRNA counts in morphologically intact cells. To detect endogenous mRNAs in single fixed cells, we optimized a single molecule fluorescent <em>in situ</em> hybridization (smFISH) protocol for <em>C. albicans</em>, which allows one to quantify the differential expression of mRNAs in yeast, pseudohyphae or hyphal cells. We quantified the expression of two mRNAs, cell cycle-controlled mRNA (<em>CLB2)</em> and a transcription regulator (<em>EFG1</em>), which show differential expression in the different morphological cell types and in different nutrient conditions. In this protocol, we described in detail the major steps of this approach: growth and fixation, hybridization, imaging, cell-segmentation and mRNA spot analysis. Raw data is provided with the protocol to favour reproducibility. This approach could benefit the molecular characterization of <em>C. albicans</em> and other filamentous fungi, pathogenic or non-pathogenic.</p> <p><strong>Data description:</strong></p> <p>This dataset consists of a FISH experiment spanning two different mRNAs, EFG1 and CLB2, and one nutrient condition, SPIDER37, in Candida albicans. For culturing, the C. albicans wildtype strain SC5314 was inoculated at 30 degrees overnight (~15 hours) in 10 mL of TSB medium in a 30 °C shaking incubator. Next, samples were diluted to a density of 10^5 cells/ mL and inoculated for 6 hours in 30 mL Spider medium at 37 °C in falcon tubes on an orbital microplate shaker. Then, samples were fixated by adding PFA to a final concentration of 4% to the medium. For hybridization, both mRNAs were hybridized independently by specific DNA oligo labelled with a Quasar670 dye to enable the visualisation of single mRNA molecules. As both genes are labelled by the same dye, these oligos were not co-applied to the same sample but to independent samples.</p> <p><strong>Microscopy</strong></p> <p>For smFISH imaging we use an Olympus BX-63 epifluorescence microscope equipped with Ultrasonic stage and UPlanApo 100x 1.35NA oil-immersion objective (Olympus). Lumencore SOLA FISH light source, a Hamamatsu ORCA-Fusion sCMOS camera (6.5 µm-pixel size) mounted using U-CMT C-Mount Adapter, and zero-pixel shift filter sets: F36-500 DAPI HC Brightline Bandpass Filter, F36-502 FITC HC BrightLine Filter, F36-542 Cy3 HC BrightLine Filter, and F36-523 Cy5 HC BrightLine Filter. Images are acquired across 61-81 optical sections (depending on the sample thickness) with a z-step size of 0.2 μm. The CellSens software (Olympus) is used for instrument control and image acquisition. For the DAPI channel 10-50 ms of exposure was used. Whilst, for the CY5 channel, used for imaging the FISH probes, 750 ms was applied. </p>
Data from: Detecting signals of chronic shedding to explain pathogen persistence: Leptospira interrogans in California sea lions
Identifying mechanisms driving pathogen persistence is a vital component of wildlife disease ecology and control. Asymptomatic, chronically infected individuals are an oft-cited potential reservoir of infection but demonstrations of the importance of chronic shedding to pathogen persistence at the population level remain scarce. Studying chronic shedding using commonly collected disease data is hampered by numerous challenges, including short-term surveillance that focuses on single epidemics and acutely ill individuals, the subtle dynamical influence of chronic shedding relative to more obvious epidemic drivers, and poor ability to differentiate between the effects of population prevalence of chronic shedding versus intensity and duration of chronic shedding in individuals. We use chronic shedding of Leptospira interrogans serovar Pomona in California sea lions (Zalophus californianus) as a case study to illustrate how these challenges can be addressed. Using leptospirosis-induced strands as a measure of disease incidence, we fit models with and without chronic shedding, and with different seasonal drivers, to determine the timescale over which chronic shedding is detectable and the interactions between chronic shedding and seasonal drivers needed to explain persistence and outbreak patterns. Chronic shedding can enable persistence of L. interrogans within the sea lion population. However, the importance of chronic shedding was only apparent when surveillance data included at least two outbreaks and the intervening inter-epidemic trough during which fadeout of transmission was most likely. Seasonal transmission, as opposed to seasonal recruitment of susceptibles, was the dominant driver of seasonality in this system, and both seasonal factors had limited impact on long-term pathogen persistence. We show that the temporal extent of surveillance data can have a dramatic impact on inferences about population processes, where the failure to identify both short- and long-term ecological drivers can have cascading impacts on understanding higher-order ecological phenomena, such as pathogen persistence.
Fig. 1 in Detection of rodent-borne parasitic pathogens of wild rats in Serdang, Selangor, Malaysia: A potential threat to human health
Fig. 1. The study area, Universiti Putra Malaysia showing the study sites circled in black.
Out of the blue: Detection of a unique highly pathogenic avian influenza virus of subtype H7N5 in Germany: Data sets on phylogenetic analyses
<div> <p><span>In June 2024, a highly pathogenic avian influenza virus of subtype H7N5 was detected in a single laying hen holding in Germany. Closest relatives of the eight viral genome segments were among recent low pathogenic (LP) viruses from Asia and Europe. No further detections of this unique virus or its presumed LP precursor have been made in poultry or wild birds.</span></p> </div>
NPIP: A Comprehensive Analysis Pipeline for Rapid Pathogen Detection in Clinical Samples Based on Nanopore Sequencing
<p>Background: Rapid and accurate pathogen detection is important for effective control of infectious diseases. However, traditional pathogen culture methods have a very long detection time, as well as high rates of false-positive and false-negative results. Third generation sequencing (TGS) technology brings the new possibility of being used as a pathogen detection method. However, the practicability of a pathogen detection report based on TGS is still lacking. There is also a lack of professional and accurate report interpretation.</p> <p>Results: Here, we report on the development of a pathogen detection and analysis tool (NPIP) based on third generation nanopore sequencing technology. We also prove the practicability of nanopore sequencing and NPIP analysis tools in emergency and clinical pathogen detection by demonstrating its use in a practical case.</p> <p>Conclusions: This platform provides an effective, convenient, and fast analysis tool for clinicians and public health personnel to more successfully apply TGS in pathogen detection.</p>
Data for: Detection of oomycete pathogens in UK peat-free growing media and implications for plant health
<p>This dataset on Zenodo accompanies the manuscript Frederickson-Matika <em>et al.</em> (2024), Detection of oomycete pathogens in UK peat-free growing media and implications for plant health.</p> <p>There are two files:</p> <ul> <li>metadata.tsv - plain text table as tab-separated variables</li> <li>raw_data.tar.gz - compressed archive of 43 paired raw FASTQ files</li> </ul> <p>This represents a subset of two complete Illumina MiSeq plates (in two dated folderes) run at the James Hutton Institute containing other environmental samples using the same protocol. Only the synthetic controls and peat-free samples are provided here.<br><br>To repeat the analysis described in the paper, first install THAPBI PICT. See <a href="https://github.com/peterjc/thapbi-pict/">https://github.com/peterjc/thapbi-pict/ </a>for instructions. At the time of the paper, v1.0.14 was the current release.</p> <p>Next, decompress the raw data into a folder of paired gzipped FASTQ files. There is no need to decompress those:</p> <pre><code> $ tar -zxvf raw_data.tar.gz<br> $ ls -1 plate_20220505/ plate_20230608/</code></pre> <p>If you wish, verify the checksums to confirm the data integrity:</p> <pre><code> $ cd plate_20220505/ $ md5sum -c MD5SUM.txt<br> $ cd ../plate_20230608/ $ md5sum -c MD5SUM.txt<br> $ cd ..</code></pre> <p>Setup output directories:</p> <pre><code><code> $ mkdir -p intermediate/ summary/</code></code></pre> <pre>Run the THAPBI PICT pipeline:</pre> <pre><code> $ thapbi_pict pipeline -m 1s3g \<br> -i plate_*/ -o summary/peat-free \<br> -y plate_*/GBL*.fastq.gz \<br> -n plate_*/GBL*.fastq.gz \<br> -s intermediate/ \<br> -t metadata.tsv -u \<br> -x 9 -c 1,2,3,4,5,6,7,8</code><br><br></pre> <p>The options here are as follows:</p> <ul> <li>-i - two input directories of paired raw FASTQ files.</li> <li>-n - negative controls used to increase the absolute abundance threshold</li> <li>-y - synthetic controls used to increase the fractional abundance threshold</li> <li>-s - optional location to store intermediate files</li> <li>-o - output stem for reports</li> <li>-t - filename for tab-separated-variable metadata</li> <li>-u - show unsequenced samples defined in the metadata</li> <li>-x - which metadata column contains Illumina FASTQ filename stems</li> <li>-c - which metadata columns to include in the report.</li> </ul> <p>This assumes the following key default settings:</p> <ul> <li>-a 100 (default absolite abundance threshold)</li> <li>-f 0.001 (default fractional abundance threshold)</li> <li>-d -(default provided ITS1 database).</li> </ul> <p>With these settings, only synthetic sequences were found in the controls, and therefore the thresholds were not automatically increased any further.</p> <p>Opening the output file summary/peat-free.ITS1.samples.1s3g.xlsx in Excel or similar should show you a table resembling Table 1 in the paper, but one row per sequencing sample, and additional columns with per-sample per-species read counts etc.</p>
Improvements Through the Use of a Rapid Multiplex PCR Enteric Pathogen Detection Kit in Children With Hematochezia
ClinicalTrials.gov study NCT03362970. IPD Sharing: NO. Countries: 1. Publications: 5.
Data from: A novel molecular toolkit for rapid detection of the pathogen and primary vector of thousand cankers disease
Open the record for dataset details and reuse information.
A method for characterizing disease emergence curves from paired pathogen detection and serology data
Open the record for dataset details and reuse information.
Data from: Detecting signals of chronic shedding to explain pathogen persistence: Leptospira interrogans in California sea lions
Open the record for dataset details and reuse information.
Data from: Imperfect pathogen detection from non-invasive skin swabs biases disease inference
Open the record for dataset details and reuse information.
Data from: Evolution of Drosophila resistance against different pathogens and infection routes entails no detectable maintenance costs
Pathogens exert a strong selective pressure on hosts, entailing host adaptation to infection. This adaptation often affects negatively other fitness-related traits. Such trade-offs may underlie the maintenance of genetic diversity for pathogen resistance. Trade-offs can be tested with experimental evolution of host populations adapting to parasites, using two approaches: (1) measuring changes in immunocompetence in relaxed-selection lines and (2) comparing life-history traits of evolved and control lines in pathogen-free environments. Here, we used both approaches to examine trade-offs in Drosophila melanogaster populations evolving for over 30 generations under infection with Drosophila C Virus or the bacterium Pseudomonas entomophila, the latter through different routes. We find that resistance is maintained after up to 30 generations of relaxed selection. Moreover, no differences in several classical life-history traits between control and evolved populations were found in pathogen-free environments, even under stresses such as desiccation, nutrient limitation, and high densities. Hence, we did not detect any maintenance costs associated with resistance to pathogens. We hypothesize that extremely high selection pressures commonly used lead to the disproportionate expression of costs relative to their actual occurrence in natural systems. Still, the maintenance of genetic variation for pathogen resistance calls for an explanation.
Image Dataset for 'AI-enabled biosensing for rapid pathogen detection: from liquid food to agricultural water'
<p>This dataset is presented in the following publication. Please cite this publication if you use the dataset.</p> <p><em>Jiyoon Yi, Nicharee Wisuthiphaet, Pranav Raja, Nitin Nitin, J. Mason Earles. (2023). AI-enabled biosensing for rapid pathogen detection: from liquid food to agricultural water. Water Research, 120258. doi: <a href="https://doi.org/10.1016/j.watres.2023.120258">10.1016/j.watres.2023.120258</a></em></p>
Supplementary material 2 from: Colombari F, Battisti A (2023) Citizen science at school increases awareness of biological invasions and contributes to the detection of exotic ambrosia beetles. In: Jactel H, Orazio C, Robinet C, Douma JC, Santini A, Battisti A, Branco M, Seehausen L, Kenis M (Eds) Conceptual and technical innovations to better manage invasions of alien pests and pathogens in forests. NeoBiota 84: 211-229. https://doi.org/10.3897/neobiota.84.95177
Slides of the lecture on 'Monitoring of insect species harmful to trees and forests' and link to educational videos
Pathogen Detection in HIV-infected Children With Non-malarial Febrile Illnesses Using Metagenomic Sequencing
ClinicalTrials.gov study NCT05085158. IPD Sharing: UNDECIDED. Countries: 1. Publications: 12.
ScienceDex guides
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These curated guides explain access requirements, typical timelines, costs, and reuse considerations for widely used research datasets.
Allen Brain Atlas
Allen Brain Atlas is an Allen Institute collection of brain map atlases, datasets, APIs, and analysis tools covering mouse, human, and non-human primate brain resources.
Annotated Behaviour and Observability Dataset (ABODe)
ABODe is a University of Edinburgh DataShare dataset for behavior classification in group-housed mice using home-cage video, identities, bounding boxes, ground-plate positions, and annotator labels.
DANDI Archive for NWB datasets
DANDI is a BRAIN Initiative archive for publishing and sharing neurophysiology data, including electrophysiology, optophysiology, and behavioral data packaged as NWB and related standards.
International Brain Laboratory public data
The International Brain Laboratory public data releases expose standardized mouse decision-making experiments, including Neuropixels recordings, widefield calcium imaging, behavior, and session metadata accessed through the ONE API.
OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.