COVID-19 PubSeq Uploading Data (part 3)

Table of Contents

1 Uploading Data

The COVID-19 PubSeq allows you to upload your SARS-Cov-2 strains to a public resource for global comparisons. A recompute of the pangenome gets triggered on upload. Read the ABOUT page for more information.

2 Step 1: Upload sequence

To upload a sequence in the web upload page hit the browse button and select the FASTA file on your local hard disk.

We start with an assembled or mapped sequence in FASTA format. The PubSeq uploader contains a QC step which checks whether it is a likely SARS-CoV-2 sequence. While PubSeq deduplicates sequences and never overwrites metadata, you may still want to check whether your data already is in the system by querying some metadata as described in Query metadata with SPARQL or by simply downloading and checking one of the files on the download page. We find GenBank MT536190.1 has not been included yet. A FASTA text file can be downloaded to your local disk and uploaded through our web upload page. Make sure the file does not include any HTML!

Note: we currently only allow FASTA uploads. In the near future we'll allow for uploading raw sequence files. This is important for creating an improved pangenome.

3 Step 2: Add metadata

The web upload page contains fields for adding metadata. Metadata is not only important for attribution, is also important for analysis. The metadata is available for queries, see Query metadata with SPARQL, and can be used to annotate variations of the virus in different ways.

A number of fields are obligatory: sample id, date, location, technology and authors. The others are optional, but it is valuable to enter them when information is available. Metadata is defined in this schema. From this schema we generate the input form. Note that optional fields have a question mark in the type. You can add metadata yourself, btw, because this is a public resource! See also Modify metadata for more information.

To get more information about a field click on the question mark on the web form. Here we add some extra information.

3.1 Obligatory fields

3.1.1 Sample ID (sampleid)

This is a string field that defines a unique sample identifier by the submitter. In addition to sampleid we also have hostid, providersampleid and submittersampleid where host is the host the sample came from, provider sample is the institution sample id and submitter is the submitting individual id. hostid is important when multiple sequences come from the same host. Make sure not to have spaces in the sampleid.

Here we add the GenBank ID MT536190.1.

3.1.2 Collection date

Estimated collection date. The GenBank page says April 6, 2020.

3.1.3 Collection location

A search on wikidata says Los Angeles is https://www.wikidata.org/entity/Q65

3.1.4 Sequencing technology

GenBank entry says Illumina, so we can fill that in

3.1.5 Authors

GenBank entry says 'Lamers,S., Nolan,D.J., Rose,R., Cross,S., Moraga Amador,D., Yang,T., Caruso,L., Navia,W., Von Borstel,L., Hui Zhou,X., Freehan,A. and Garcia-Diaz,J.', so we can fill that in.

3.2 Optional fields

All other fields are optional. But let's see what we can add.

3.2.1 Host information

Sadly, not much is known about the host from GenBank. A little sleuthing renders an interesting paper by some of the authors titled SARS-CoV-2 is consistent across multiple samples and methodologies which dates after the sample, but has no reference other than that the raw data came from the SRA database, so it probably does not describe this particular sample. We don't know what this strain of SARS-Cov-2 did to the person and what the person was like (say age group).

3.2.2 Collecting institution

We can fill that in.

3.2.3 Specimen source

We have that: nasopharyngeal swab

3.2.4 Source database accession

Genbank which is http://identifiers.org/insdc/MT536190.1#sequence. Note we plug in our own identifier MT536190.1.

3.2.5 Strain name

SARS-CoV-2/human/USA/LA-BIE-070/2020

4 Step 3: Submit to COVID-19 PubSeq

Once you have the sequence and the metadata together, hit the 'Add to Pangenome' button. The data will be checked, submitted and the workflows should kick in!

4.1 Trouble shooting

We got an error saying: {"stem": "http://www.wikidata.org/entity/",… which means that our location field was not formed correctly! After fixing it to look like http://www.wikidata.org/entity/Q65 (note http instead on https and entity instead of wiki) the submission went through. Reload the page (it won't empty the fields) to re-enable the submit button.

5 Step 4: Check output

The current pipeline takes 5.5 hours to complete! Once it completes the updated data can be checked on the DOWNLOAD page. After completion of above output this SPARQL query shows some of the metadata we put in.

6 Bulk sequence uploader

Above steps require a manual upload of one sequence with metadata. What if you have a number of sequences you want to upload in bulk? For this we have a command line version of the uploader that can directly submit to COVID-19 PubSeq. It accepts a FASTA sequence file an associated metadata in YAML format. The YAML matches the web form and gets validated from the same schema looks. The YAML that you need to create/generate for your samples looks like

id: placeholder

host:
    host_id: XX1
    host_species: http://purl.obolibrary.org/obo/NCBITaxon_9606
    host_sex: http://purl.obolibrary.org/obo/PATO_0000384
    host_age: 20
    host_age_unit: http://purl.obolibrary.org/obo/UO_0000036
    host_health_status: http://purl.obolibrary.org/obo/NCIT_C25269
    host_treatment: Process in which the act is intended to modify or alter host status (Compounds)
    host_vaccination: [vaccines1,vaccine2]
    ethnicity: http://purl.obolibrary.org/obo/HANCESTRO_0010
    additional_host_information: Optional free text field for additional information

sample:
    sample_id: Id of the sample as defined by the submitter
    collector_name: Name of the person that took the sample
    collecting_institution: Institute that was responsible of sampling
    specimen_source: [http://purl.obolibrary.org/obo/NCIT_C155831,http://purl.obolibrary.org/obo/NCIT_C155835]
    collection_date: "2020-01-01"
    collection_location: http://www.wikidata.org/entity/Q148
    sample_storage_conditions: frozen specimen
    source_database_accession: [http://identifiers.org/insdc/LC522350.1#sequence]
    additional_collection_information: Optional free text field for additional information

virus:
    virus_species: http://purl.obolibrary.org/obo/NCBITaxon_2697049
    virus_strain: SARS-CoV-2/human/CHN/HS_8/2020

technology:
    sample_sequencing_technology: [http://www.ebi.ac.uk/efo/EFO_0009173,http://www.ebi.ac.uk/efo/EFO_0009173]
    sequence_assembly_method: Protocol used for assembly
    sequencing_coverage: [70.0, 100.0]
    additional_technology_information: Optional free text field for additional information

submitter:
    authors: [John Doe, Joe Boe, Jonny Oe]
    submitter_name: [John Doe]
    submitter_address: John Doe's address
    originating_lab: John Doe kitchen
    lab_address: John Doe's address
    provider_sample_id: XXX1
    submitter_sample_id: XXX2
    publication: PMID00001113
    submitter_orcid: [https://orcid.org/0000-0000-0000-0000,https://orcid.org/0000-0000-0000-0001]
    additional_submitter_information: Optional free text field for additional information

6.1 Run the uploader (CLI)

Installing with pip you should be able to run

bh20sequploader sequence.fasta metadata.yaml

Alternatively the script can be installed from github. Run on the command line

python3 bh20sequploader/main.py example/sequence.fasta example/maximum_metadata_example.yaml

after installing dependencies (also described in INSTALL with the GNU Guix package manager).

The web interface using this exact same script so it should just work (TM).

6.2 Example: uploading bulk GenBank sequences

We also use above script to bulk upload GenBank sequences with a FASTA and YAML extractor specific for GenBank. This means that the steps we took above for uploading a GenBank sequence are already automated.


Created by Pjotr Prins (pjotr.public768 at thebird 'dot' nl) using Emacs org-mode and a healthy dose of Lisp!
Modified 2020-08-22 Sat 07:43
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