A negative PCR comparison needs a condition ledger
The title describing improved TaqMan real-time PCR assays for pathogenic Leptospira leaves an important replication question open. If a laboratory does not recover the reported performance, the result is informative only after separating assay failure from changed conditions. A replication record should identify what remained fixed across primer and probe sequences, reagent lots, extraction method, specimen matrix, target concentrations, thermocycler settings, threshold rules, and reference classification. Methodological work on reproducibility treats laboratory conditions, batches, personnel, and protocols as sources of heterogeneity rather than automatic evidence against the original finding. What was held constant, and which single planned variation would test whether the apparent failure marks a boundary condition?
