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Z-score background correction

Virasign can compute a background-corrected Z-score per reported virus using water controls (e.g. negative controls). This mirrors a common idea in metagenomic reporting: quantify whether a signal is unusually high compared to background contamination.

Why use a Z-score?

Some taxa show up at low levels in many runs due to:

  • reagent / extraction contamination
  • index hopping / low-level carryover
  • laboratory environment background

By comparing each virus to your water controls in the same run, the Z-score answers:

“Is this virus higher than what we typically see in water controls?”

When Virasign computes it

  • Z-scores are computed only when ≥2 water controls are available.
  • Water controls are auto-detected when the sample name contains water, h2o, or h20 (case-insensitive).
  • You can override auto-detection with exact input FASTQ paths:
virasign -i input_dir --zscore-controls /path/to/water1.fastq.gz,/path/to/water2.fastq.gz

When --zscore-controls is provided, Virasign does not use auto-detection (so you can exclude some “H2O_*” samples intentionally).

What signal is used

Virasign computes the Z-score using the per-hit remapped mapped_reads (the same value reported in the final per-sample JSON).

Formula

Virasign computes, for each virus, a Z-score as the number of standard deviations that the sample’s log-transformed mapped_reads signal is above/below the mean of the log-transformed mapped_reads values in the selected water controls.

This follows the same background-correction idea used by CZ ID / IDseq background models. For more information, see:

  • IDseq paper (GigaScience, 2020): https://ncbi.nlm.nih.gov/pmc/articles/PMC7566497/
  • CZ ID workflows wiki: https://github.com/chanzuckerberg/czid-workflows/wiki

How to interpret Z-scores

Z-scores are “standard deviation units” above/below background. Practical interpretation depends on context and other evidence (coverage breadth/depth, identity, NOGR, etc.), but the table below is a useful starting point.

Z-score SD interpretation Practical interpretation
-3 3 SD below mean Below background
-1 1 SD below mean Background-like
0 At mean Background-like
1 1 SD above mean Slightly above background
2 2 SD above mean Above background
3 3 SD above mean Strongly above background
10 10 SD above mean Strongly above background
50 Extreme outlier Strongly above background
100 Extreme outlier typically only in non-water samples

Where it appears in outputs

  • Per-hit fields in *_final_selected_references.json:
    • zscore: the computed value
    • zscore_controls: list of water samples used for the background model
  • results_summary_*.html and results_summary_*.csv include a Z-score column.