Consolidated Comparison of Immunoassay Performance Specifications
A current comparison of Global Performance Specifications for Immunoassay Assays.
GAMECHANGE: AN ON-DEMAND WEBINAR ON CLIA
Consolidated Comparison of Immunoassay (and Tumor Marker) Performance Specifications
Sten Westgard, MS
Updated Feburary 7, 2024 with latest EFLM specifications, RCPA specifications
Updated March 22, 2023 with addition of Brazilian goals, removal of RCPA goals
Updated July 24, 2022 with proposed CLIA 2024 proficiency testing criteria
Updated June 14, 2022 with EFLM minimum quality performance specifications
Updated February 24, 2022 with 2022 critically evaluated tumor marker goals[g] and thyroid biomarkers[h]
Updated February 17, 2022 with 2021 RIQAS state of the art goals
Updated January 21, 2022
Extending our consolidation and comparison of performance specifications, started in chemistry, into immunoassays, as well as endocrinology and cancer markers and drugs of abuse. Traditionally, immunoassays have been given fewer goals, and those that are given are generally more permissive. The CLIA targets and other EQA/PT goals are often in sharp contrast to EFLM biological database goals, showing either a gap in the capability of instrumentation, or possibly a lack of will in regulation.
| Routine Immunoassay Performance Specifications (allowable total analytical error, TEa) | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Test or Analyte | Recommended Choice | CLIA | CLIA "2024" | Riqas 2020 SOTA | 2014 Desirable "Ricos goals" |
EFLM/ |
Rilibak | 2022 RCPA ALP | Brazil | 2020 "Spanish Min."[k] |
| Aldosterone | ± 42.8%[f] ± 64.2%[i] |
± 24 pmol/L; 15% @ 160 pmol/L |
||||||||
| Alpha-Fetoprotein (AFP) | ± 20.0% | -- | TV ± 20% or positive or negative | ±13.3% |
± 21.9% |
± 34.83% ± 17.6%[g] ± 26.5%[i] |
± 24% | ± 2 kIU/L; 12% @ 17 kIU/L |
± 18% | |
| Beta-2 microglobulin | -- | -- | ± 6.4%[f] ± 9.7%[i] |
± 0.2 mg/L; 10% @ 2.0 mg/L |
</td | ± 14% | ||||
| Human Chorionic Gonadotropin (BhCG) | ± 18% or ± 3 mIU/mL (greater) |
-- | ± 18% or ± 3 mIU/mL or positive or negative |
± 30% | ± 1 IU/L; 10% @ 10 IU/L |
± 20% | ± 14% | |||
| CA 125 | ± 20.0% | -- | ± 20% | ±12.9% | ± 35.4% |
± 13.9%[f] |
± 6 kU/L; 12% @ 50 kU/L |
± 11% | ||
| CA 15-3 | ± 20.8% | -- | -- | ±14.3% | ± 20.8% | ±12.9%[g] | ± 24% | ± 3 kU/L; 10% @ 30 kU/L |
± 13% | |
| CA 19-9 | ± 26.9% | -- | -- | ±16.5% | ± 46.0% | ± 17.9%[f] ± 18.0%[g] ± 26.9%[i] |
± 6 kU/L; 15% @ 40 kU/L |
± 13% | ||
| CA 72-4 | ± 70.2% | -- | -- | ± 70.2%[f,g] ± 105.4%[i] |
-- | ± 11% | ||||
| Carbamazepine | ± 1.0 mcg/dL or ± 20.0% |
± 25.0% | ± 1.0 mcg/dL or ± 20.0% |
±13.7% | -- | -- | ± 20% | ± 0.5 mg/L; 10% @ 4.7 mg/L |
± 0.5 mg/L or 10% | |
| Calcitonin | ± 27.5% | -- | -- |
± 27.5%[f, h] |
± 2 ng/L; 10% @ 20 ng/L |
|||||
| Carcinogenic Embryonic Antigen (CEA) | ± 15% or ± 1 ng/dL | -- | ± 15% or ± 1 ng/dL | ±11.3% | ± 24.7% | ± 26.94% ± 20.5%[f] ± 30.8%[i] |
± 24% | ± 0.6 ug/L; 12% @ 5.0 ug/L |
± 16.0% | |
| Ceruloplasmin | ± 8.1%[f] ± 12.2%[i] |
-- | ± 12% | |||||||
| CK-MB | ± 25.0% | ± 25.0% | ± 25.0% | ± 16.5% | -- | -- | ± 15% | |||
| Cobalamin | ± 15.4%[f] ± 23.0%[i] |
-- | ± 14% | |||||||
| Cortisol | ± 20.0% | ± 25.0% | ± 20.0% | ±13.5% | ± 22.8% | ± 22.6%[f] ± 33.9%[i] |
± 30% | ± 15 nmol/L; 15% @ 100 nmol/L |
± 25% | ± 19% |
| C-peptide | ± 20.8% | -- | -- | ±14.8% | ± 20.8% | -- | ± 0.15 nmol/L; 12% @ 1.25 nmol/L |
± 12% | ||
| Cystatin-C | ± 9.7% | -- | -- | ± 7.6% | ± 5.4%[f] ± 8.0%[i] |
-- | ||||
| DHT | -- | -- | ± 19.3%[f] ± 29.0%[i] |
-- | ||||||
| Digoxin | ± 15% or ± 0.2 ng/mL | ± 0.2 ng/mL or 20% | ± 15% or ± 0.2 ng/mL | ±14.9% | -- | ± 30% | ± 0.2 ug/L; 10% @ 1.6 ug/L |
± 21% | ||
| Estradiol | ± 30% | -- | ± 30% | ±13.7% | ± 26.86% | ± 18.3%[f] ± 27.4%[i] |
± 35% | ± 25 pmol/LI; 25% @ 100 pmol/LI |
± 20% | ± 21% |
| Ferritin | ± 20.0% | -- | ± 20% | ±13.1% | ± 16.9% | ± 13.9%[f] ± 20.8%[i] |
± 25% | ± 4.0 ug/L; 15% @ 27.0 ug/L |
± 18% | |
| Folate | ± 1 ng/mL or 30% | ± 1 ng/mL or 30% | ±23.9% | ± 39% | ± 15.5%[f] ± 23.3%[i] |
± 1.5 nmol/L; 25% @ 6.0 nmol/L |
± 20% | |||
| Follicle Stimulating Hormone (FSH) | ± 2 IU/L or 18% | ± 2 IU/L or 18% | ±11.4% | ± 21.2% | ± 17.6%[f] ± 26.4%[i] |
± 21% | ± 1.0 IU/L; 10% @ 10.0 IU/L |
± 20% | ± 14% | |
| Free kappa light chain | ±22.8% | ±22.8% | ± 8.0% | ± 8.8%[f] ± 13.2%[i] |
-- | |||||
| Free lamba light chain | ±22.6% | ±22.6% | ± 8.6% | ± 10.3%[f] ± 15.4%[i] |
-- | |||||
| Fructosamine | ± 3.6%[f] ± 5.4%[i] |
-- | ||||||||
| Gentamicin | ± 25% | ± 25% | ± 25% | ±13.3% | -- | ± 0.2 mg/L; 10% @ 2.0 mg/L |
||||
| Glycine | ± 19.9%[f] ± 29.8%[i] |
-- | ||||||||
| Homocysteine | ± 15.4% | ±14.3% | ± 15.4% | -- | ± 1.5 umol/L; 10% @ 15.0 umol/L |
± 17% | ||||
| IGF-1 | ± 14.9%[f] ± 22.4%[i] |
± 3 nmol/L; 12% @ 25 nmol/L |
||||||||
| IL6 | ± 39.0%[f] ± 58.4%[i] |
-- | ||||||||
| Insulin | ± 32.9% | ±15.3% | ± 32.9% | ± 31.5%[f] ± 47.2%[i] |
± 0.6 mU/L; 12% @ 5.0 mU/L |
± 38% | ||||
| Luteinizing Hormone (LH) | ± 20.0% | ± 20% | ±12.1% | ± 27.92% | ± 29.1%[f] ± 43.6%[i] |
± 1.5 IU/L; 15% @ 10.0 IU/L |
± 25% | |||
| Parathyroid Hormone (PTH I-84) | ± 30.0% | ± 30% | ±21.2% | ± 30.2% | ± 19.6%[f] ± 29.4%[i] |
± 1.0 pmol/L; 12% @ 8.0 pmol/L |
± 23% | |||
| Phenobarbital | ± 15% or ± 2 mcg/mL | ± 20% | ± 15% or ± 2 mcg/mL | ±13.7% | -- | ± 20% | ± 0.7 mg/L; 10% @ 7.0 mg/L |
± 15% | ||
| Phenytoin | ± 2 mcg/dL or 15% | ± 25% | ± 2 mcg/dL or 15% | ±14.6% | -- | ± 20% | ± 0.8 mg/L; 10% @ 7.6 mg/L |
± 13% | ||
| Progesterone | ± 25% | ± 25% | ± 25% | ±15.6% | ± 26.2% [f] ± 39.3% [i] |
± 35% | ± 2.0 nmol/L; 15% @ 10 nmol/L |
± 30% | ± 22% | |
| Prolactin | ± 20% | ± 20% | ±17.1% | ± 29.4% | ± 56.7% [f] ± 85.1% [i] |
± 40 mIU/L; 10% @ 400 mIU/L |
± 20% | ± 22% | ||
| Prostate Specific Antigen (PSA) including free (f), conjugated (c), and total (t) |
± 0.2 ng/mL or 20% | ± 0.2 ng/mL or 20% | ±11.6% | ± 33.6% | c±21.9%[g] f±17.5%[g] t±16.2%[g] t±16.2%[f] t±24.4%[i] c±21.9%[f] c±32.8%[i] f±17.5%[f] f±26.3%[i] |
± 25% | total ± 0.4 ug/L; 8% @ 5.0 ug/L |
± 15%(t) ± 13%(f) |
||
| Renin | ± 37.7%[f] ± 56.5%[i] |
-- | ||||||||
| S100B | ± 17.0%[f] ± 25.5%[i] |
-- | ||||||||
| Sex Hormone Binding Globulin (SHBG) | ± 26.1% | ±13.7% | ± 20.4% | ± 19.3%[f] ± 28.9%[i] |
± 6 nmol/L; 12% @ 50 nmol/L |
|||||
| sTFr | ± 10.8%[f] ± 16.2%[i] |
-- | ||||||||
| ST2 | ± 19.3%[f] ± 29.0%[i] |
-- | ||||||||
| Testosterone, including free (f) |
± 20 ng/dL or 30% | ± 20 ng/dL or 30% | ±15.5% | ± 13.6% | ± 18.8%[f] ± 28.2%[i] f± 67.2%[f] f± 100.9%[i] |
± 35% | ± 0.4 nmol/L; 15% @ 2.7 nmol/L |
± 22% | ||
| Theophylline | ± 20% | ± 25% | ± 20% | ±15.1% | -- | ± 24% | ± 0.5 mg/L; 10% @ 5.4 mg/L |
|||
| Thyroglobuin | ± 29.1%[f] ± 43.7%[i] |
± 0.2 ug/L; 12% @ 1.7 ug/L |
± 17% | |||||||
| Thyroid Stimulating Hormone (TSH) | ± 2 mIU/L or 20% | ± 2 mIU/L or 20% | ±11.2% | ± 23.7% |
± 24.8%[f,h] |
± 24% | ± 0.1 mU/L; 20% @ 0.5 mU/L |
± 20% | ± 15% | |
| Thyroxine, Free (FT4) | ± 20.0% | ± 0.3 ng/dL or 15% | ±11.9% | ± 6.2%[h] ± 6.3%[f] ± 9.4%[i] |
± 20% | ± 1.5 pmol/L; 12% @ 12 pmol/L |
± 20% | ± 16% | ||
| Thyroxine, Total (TT4) | ± 1.0 mcg/dL or 20% |
± 1.0 mcg/dL or 20% | ± 1.0 mcg/dL or 20% |
±14.5% | ± 7% | ± 8.6%[f] ± 13.0%[i] |
± 24% | ± 12 nmol/L; 10% @ 120 nmol/L |
± 1 ug/dL or 16% | ± 20% |
| TNF | ± 23.2%[f] ± 34.8%[i] |
-- | ||||||||
| TRAP | ± 13.2%[f] ± 19.2%[i] |
-- | ||||||||
| Triiodothyronine, Free (FT3) | ± 20.0% | ±13.3% | ± 11.3% | ± 6.6%[f,h] ± 9.9%[i] |
± 20% | ± 0.7 pmol/L; 20% @ 3.5 pmol/L |
± 35% | ± 12% | ||
| Triiodothyronine, Total (TT3) | ± 30.0% | ± 30.0% | ±16.4% | ± 9.22% | ± 8.3%[f] ± 12.4%[i] |
± 24% | ± 0.2 nmol/L; 15% @ 1.3 nmol/L |
± 20% | ± 20% | |
| Troponin I | ± 0.9 ng/mL or 30% | ± 0.9 ng/mL or 30% | ±19.1% | ± 27.9% | hs±20.0%[f] hs± 29.9%[i] |
± 33% | ± 0.002 ug/L; 20% @ 0.01 ug/L |
± 21% | ||
| Troponin T | ± 0.2 ng/mL or 30% | ± 0.2 ng/mL or 30% | ± 48.9% | hs± 17.6%[f] hs± 26.5%[i] |
± 33% | ± 0.01 ug/L; 20% @ 0.05 ug/L |
± 22% | |||
| Tyrosine | ± 20.7%[f] ± 31.0%[i] |
-- | ||||||||
| Valproic Acid | ± 25% | ± 25% | ± 25% | ±12.1% | -- | ± 20% | ± 4.0 mg/L; 10% @ 0.01 ug/L |
|||
| Vancomycin | ± 2.0 mcg/dL or 15% | ± 2.0 mcg/dL or 15% | ±16.2% | -- | ± 18% | ± 2.0 mg/L; 10% @ 20.3 mg/L |
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| Vitamin A | ± 10.5%[f] ± 15.6%[i] |
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| Vitamin B1 | ± 7.4%[f] ± 11.1%[i] |
|||||||||
| Vitamin E | ± 10.5%[f] ± 15.8%[i] |
|||||||||
| Vitamin K | ± 45.9%[f] ± 68.8%[i] |
|||||||||
| Vitamin B12 | ± 25% or ± 30 pg/mL | ± 30% | ± 25% or ± 30 pg/mL | ±17.3% | ± 30% | -- | ||||
Not all analytes are listed in this table. But this gives an idea of the global range of goals.
Here are the references for the EuBIVAS studies:
a. Biological Variation Estimates Obtained from 91 Healthy Study Participants for 9 Enzymes in Serum, Carobene A, Roraas T, Solvik UO, et al, Clin Chem 63:6. 2018.
b. EuBIVAS: Within- and Between-Subject Biological Variation Data for Electrolytes, Lipids, Urea, Uric Acid, Total Protein, Total Bilirubin, Direct Bilirubin, and Glucose, Aarsand AK, Diaz-Garzon J, Fernandez-CAlle P et al, Clin Chem 64:9 2017
c. The EuBIVAS Project: Within- and Between-Subject Biological Variation Data for Serum Creatinine Using Enzymatic and Alkaline Picrate Methods and Implications for Monitoring, Carobene A, Marino I, Coskun A et al, Clin Chem 63:9. 2017 Estimates from Enzymatic methods used.
d. Biological variation data for lipid cardiovascular risk assessment biomarkers. A systematic review applying the biological variation data critical appraisal checklist (BIVAC), Diaz-Garzon J, Fernandez-CAlle P, Minchinel J et al Clin Chim Acta 495 (August 2019) 467-475.
e. Systematic review of the biological variation data for diabetes related analytes, Gonzalez-Lao E, Corte Z, Simon M et al, Clin Chim Acta 488 (January 2019) 61-67.
f. EFLM database, originally accessed 1/13/2020. most recently updated 2/7/2024 https://biologicalvariation.eu/meta_calculations
g. Critical review and meta-analysis of biological variation estimates for tumor markers, Marques-Garcia F, Boned B, Gonzalez-Lao E et al. Clin Chem Lab Med 2022 https://doi.org/10.1515/cclm-2021-0725
h. European Biological Variation Study (EuBIVAS): within- and between-subject biological variation estimates for serum thyroid biomarkers based on weekly samplings from 91 healthy participants. Bottani M, Aarsand AK, Banfi G et al. Clin Chem Lab Med 2021 https://doi.org/10.1515/cclm-2020-1885
i. EFLM database, originally accessed 6/13/2022. most recently updated 2/7/2024 with MINIMUM specification recommendations https://biologicalvariation.eu/meta_calculations
j. CLIA 2024 proficiency testing criteria, quoted from the Federal Register Vol 87, No 131, July 11, 2022. Full listing at https://www.westgard.com/2024-clia-requirements.htm
k. Minimum analytical quality consensus specifications in Spain. Comparison with previous, mandatory and biological variability values. Enrique Prada de Medio, Angel Mlonia Borras, Jose Alcaraz Quiles et al. Rev Med Lab 2020;1(3):93-107.
[Thanks to Randox RIQAS for sharing their State of the Art (SOTA) goals from their monthly EQA program]
[Special thanks to Vladimir Procenko, for an informal English translation of the Russia GOST goals, of which Annex A is listed here, which represents the maximum allowable analytical error for a single measurement.]