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Analytical Performance Validation: LOD, LOQ, CV%, and Linearity

1. Why Analytical Validation Matters

Analytical validation establishes that an assay measures what it claims, reliably, across its reportable range. For veterinary POCT, this is the foundation of every clinical claim — before a test can detect disease, it must first be shown to detect its analyte precisely and accurately.

This document defines the standard analytical parameters Migibio reports for every assay, and the methodology behind them.

2. Core Parameters

Parameter Definition What it proves
LOD Limit of Detection — lowest analyte concentration distinguishable from blank Analytical sensitivity
LOQ Limit of Quantitation — lowest concentration measurable with acceptable precision Lower reporting limit
Precision (CV%) Coefficient of variation — repeatability of measurement Reliability
Linearity Range over which signal is proportional to concentration Reportable range
Accuracy Closeness to a reference method (bias) Trueness
Interference Effect of matrix components (hemolysis, lipemia) Robustness

3. Limit of Detection (LOD)

Method: measure a blank (zero-analyte) sample 20 times; LOD = mean blank + 3 SD.

LOD = mean(blank) + 3 × SD(blank)
Parameter FICT Typical
Blank replicates 20
LOD 0.1–1.0 ng/ml (assay-dependent)
Confidence 95% (3 SD)

4. Limit of Quantitation (LOQ)

Method: LOQ is the lowest concentration measurable with CV% ≤ 20% (or the concentration at mean blank + 10 SD).

LOQ = mean(blank) + 10 × SD(blank)   (≈ where CV% = 20%)
Parameter FICT Typical
CV% at LOQ ≤ 20%
LOQ 0.5–5.0 ng/ml

5. Precision (CV%)

Two components, both reported:

Type Method Migibio Target
Intra-assay (repeatability) Same run, same sample, 20 replicates < 8%
Inter-assay (reproducibility) Different runs/days/operators, 20 runs < 10%
CV% = (SD / mean) × 100

6. Linearity & Reportable Range

Method: serially dilute a high-concentration sample; plot measured vs expected; linearity is the range where correlation r² ≥ 0.99.

Parameter FICT Typical
Linear range 3–4 orders of magnitude
r² (linear regression) ≥ 0.99

7. Accuracy (Bias vs Reference)

Method: compare against a reference method (e.g., ELISA) using a Bland-Altman plot and Passing-Bablok regression.

Metric FICT Typical
Correlation (r) vs ELISA > 0.95
Mean bias < 10%
Slope (Passing-Bablok) 0.9–1.1

8. Interference & Matrix Effects

Interferent Tested Concentration Acceptance
Hemoglobin (hemolysis) up to 5 g/L Bias < 10%
Triglycerides (lipemia) up to 10 g/L Bias < 10%
Bilirubin (icterus) up to 0.2 g/L Bias < 10%
Rheumatoid factor clinical range No false positive

9. Summary — Migibio Standard Validation Panel

Every Migibio assay ships with a validation report covering:

  1. LOD & LOQ
  2. Intra- & inter-assay precision (CV%)
  3. Linearity & reportable range
  4. Accuracy vs reference method (correlation + bias)
  5. Interference & cross-reactivity
  6. Lot-to-lot consistency
  7. Sample stability (serum/plasma/whole blood)

For per-assay performance numbers, see the Data Hub. For assay-specific validation reports, see Technical Reports.

Authored by: Migibio Clinical & Scientific Affairs, Guangzhou Magic Biotech Co., Ltd.

Reviewed by: Migibio R&D Quality Committee

Last updated: 2026-08-13

Disclosure: Migibio (Guangzhou Magic Biotech Co., Ltd.) is the manufacturer of the FIA680/FIA880 analyzers and FICT reagents referenced in this content. See our Editorial & Review Policy.

Contact: Martin.Wong  ·  Phone (WhatsApp): +86 13323237275  ·  Email: martinwang2024@gmail.com
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