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Competing-risk degradation evaluation: 2026-07-20

This stage tested two frozen alternatives to the original ONT direct-RNA degradation kernel: internally learned scalar correction weights and an observation model that separates degradation from technical truncation.

Rejected: alignment-predictive scalar calibration

The original intact/degraded model was evaluated at correction weights 0, 0.1, 0.25, 0.5, 0.75, 1. Cross-fitted ambiguous reads were scored by the agreement of corrected endpoint probabilities with their independent alignment score probabilities. A conservative prior favored zero and likelihood evidence was capped at 50 effective reads.

This successfully detected LongBench overcorrection. Learned weights were 0.03--0.07 in the four active 50,000-read libraries, and HCC827 at 250,000 reads improved from CCC 0.558229 to 0.562535, essentially equal to adaptive (0.562512). It failed the degradation-trajectory gate, however:

Trajectory Old auto Pearson Predictive-weight Pearson
TS10 9.8→7.7 0.96549 0.95439
TS12 9.9→7.2 0.94790 0.91991
TS12 9.9→7.3 0.93919 0.91205

The selector assigned the degradation series the same small weights as LongBench, collapsing robustness almost to adaptive. Alignment agreement is therefore not an adequate single-sample proxy for quantification benefit. The selector was removed rather than retained as unused runtime overhead.

Accepted: three-component competing-risk model

The accepted model represents a 3'-anchored read as one of:

  1. intact, concentrated in the full-length 5' endpoint bin;
  2. technical truncation, a length-invariant Beta(2,1) retained fraction; or
  3. degradation, a truncated exponential whose rate scales with physical transcript length.

Mixture weights are estimated continuously by cross-fitted MAP-EM with a weak Dirichlet(5,2,2) prior. Hazard is selected on the frozen regularized grid. The correction divides only the degradation likelihood ratio relative to the intact/technical mixture, rather than removing the entire observed endpoint profile. Its strength remains learned from degradation-versus-null evidence, the degraded mixture fraction, and each read's degradation posterior. Existing adaptive reliability and Bayes-factor safeguards remain in force.

Across LongBench dRNA, the model estimates 7.6--20.1% degraded molecules and 20.0--26.5% technical truncation at 50,000 reads. These components are separated by their different transcript-length dependence rather than endpoint shape alone.

Results

Degradation robustness

Trajectory None Pearson Old auto Pearson Competing-risk Pearson Competing-risk JSD Competing-risk L1
TS10 9.8→7.7 0.96623 0.96549 0.96500 0.15519 0.66866
TS12 9.9→7.2 0.94682 0.94790 0.94876 0.14290 0.64831
TS12 9.9→7.3 0.93781 0.93919 0.94013 0.14257 0.63269

The new model preserves the original auto robustness and slightly improves both TS12 comparisons.

LongBench matched-Illumina comparator

At 50,000 reads, competing-risk auto is within 0.00027 CCC of adaptive or better in all eight cell lines, while every cell line remains above none. It improves over the old auto in each library where old degradation correction was active. At 250,000 HCC827 reads:

Model CCC Pearson RMSE
adaptive 0.562512 0.739364 6869.94
old auto 0.558229 0.735039 6919.07
competing-risk auto 0.562581 0.739437 6869.19

Frozen truth

Dataset CCC MARD RMSE
large synthetic dRNA 0.996926 0.143267 24.0920
SIRV E0 dRNA 0.671364 0.154332 0.493293
source-labelled dRNA 0.983387 0.0001062 0.011939

These are effectively equal to the frozen adaptive optima. The model assigns only 0.69% degraded fraction to SIRV and 0.22% to the nearly full-length source-labelled set, making correction negligible without a manually selected mode.

Runtime and decision

Caching component probabilities during mixture EM reduced coverage construction on the 1.36-million-read synthetic benchmark from 11.3 s during development to 2.66 s; EM took 17.45 s. This is also below the previous two-component auto measurement of 4.49 s. The cached and uncached implementations produce the same fitted parameters and estimates.

Retain the three-component constant-hazard model. Bounded beta and piecewise challengers were subsequently tested; neither displaced the incumbent. See the challenger study. Gene-random effects and joint abundance/degradation inference remain unjustified.

Artifacts are under oarfish-evaluation-data/degradation/three-component* and oarfish-evaluation-data/longbench/three-component*.