Comparison of two commercial enzyme-linked immunosorbent assays for cerebrospinal fluid measurement of amyloid β1–42 and total tau

Mirjana Babić 1 , Željka Vogrinc 2 , Andrea Diana 3 , Nataša Klepac 4 , Fran Borovečki 4 , Patrick Hof 5 , and Goran Šimić 1
  • 1 Department of Neuroscience, Croatian Institute for Brain Research, University of Zagreb Medical School, Šalata 12, 10000, Zagreb, Croatia
  • 2 Laboratory for Neurobiochemistry, Department of Laboratory Diagnostics, University Hospital Centre Zagreb, Kišpatićeva 12, 10000, Zagreb, Croatia
  • 3 Laboratory of Neurogenesis and Neuropoiesis, Department of Biomedical Sciences, University of Cagliari, Città Universitaria di Monserrato, 09042, Monserrato (Cagliari), Italy
  • 4 Department for Functional Genomics, Center for Translational and Clinical Research, University of Zagreb Medical School, University Hospital Center Zagreb, Šalata 2, 10000, Zagreb, Croatia
  • 5 Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA


Amyloid β1–42 (Aβ1–42), total tau (t-tau), and phosphorylated tau (p-tau) are the main cerebrospinal fluid (CSF) biomarkers for early diagnosis of Alzheimer’s disease (AD). Detection of AD is critically important in view of the growing number of potential new drugs that may influence the course of the disease in its early phases. However, cut-off levels for these CSF biomarkers have not yet been established. Variability in absolute concentrations of AD biomarkers is high among studies and significant differences were noticed even within the same datasets. Variability in biomarkers levels in these assays may be due to many aspects of operating procedures. Standardization of pre-analytical and analytical procedures in collection, treatment, and storage of CSF samples is crucial because differences in sample handling can drastically influence results. Multicenter studies showed that usage of ELISA kits from different manufacturers also affects outcome. So far only very few studies tested the efficiency of ELISA kits produced by different vendors. In this study, the performance of Innogenetics (Gent, Belgium) and Invitrogen (Camarillo, CA, USA) ELISA kits for t-tau and Aβ1–42 was tested. Passing-Bablok analysis showed significant differences between Invitrogen and Innogenetics ELISA methods, making it impossible to use them interchangeably.

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  • [1] Ferri C.P., Prince M., Brayne C., Brodaty H., Fratiglioni L., Ganguli M., et al., Global prevalence of dementia: a Delphi consensus study, Lancet, 2005, 366, 2112–2117

  • [2] Cummings J.L., Khachaturian Z.S., Definitions and diagnostic criteria, In: Gauthier S. (ed.) Clinical diagnosis and management of Alzheimer’s disease, 2nd ed., Martin Dunitz, London, 2002, 3–13

  • [3] Wiltfang J., Esselman H., Maler J.M., Bleich S., Huther G., Kornhuber J., Molecular biology of Alzheimer’s dementia and its clinical relevance to early diagnosis and new therapeutic strategies, Gerontology, 2001, 47, 65–71

  • [4] Hansson O., Zetterberg H., Buchhave P., Londos E., Blennow K., Minthon L., Association between CSF biomarkers and incipient Alzheimer’s disease in patients with mild cognitive impairment: a follow-up study, Lancet Neurol., 2006, 5, 228–234

  • [5] Mattsson N., Zetterberg H., Hansson O., Andreasen N., Parnetti L., Jonsson M., et al., CSF biomarkers and incipient Alzheimer disease in patients with mild cognitive impairment, J. Am. Med. Assoc., 2009, 302, 385–393

  • [6] Šimić G., Boban M., Šarac H., Grbić K., Hof P.R., Hamann C., et al., CSF tau proteins in evaluation of patients with suspected dementia, Neurodegen. Dis., 2007, 4, 135–136

  • [7] Šimić G., Boban M., Hof P.R., Cerebrospinal fluid phosphorylated tau proteins as predictors of Alzheimer’s disease in subjects with mild cognitive impairment, Period. Biol., 2008, 110, 27–30

  • [8] Boban M., Grbić K., Mladinov M., Hof P.R., Süβmair C., Ackl N., et al., Cerebrospinal fluid markers in differential diagnosis of Alzheimer’s disease and vascular dementia, Coll. Antropol., 2008, 32, 31–36

  • [9] Boban M., Šarac H., Mimica N., Mladinov M., Süβmair C., Ackl N., et al., CSF tau proteins in differential diagnosis of dementia, Transl. Neurosci., 2010, 1, 43–48

  • [10] Spies P.E., Slats D., Sjögren J.M., Kremer B.P., Verhey F.R., Rikkert M.G., et al., The cerebrospinal fluid amyloid beta42/40 ratio in the differentiation of Alzheimer’s disease from non-Alzheimer’s dementia, Curr. Alzheimer Res., 2010, 7, 470–476

  • [11] Motter R., Vigo-Pelfrey C., Kholodenko D., Barbour R., Johnson-Wood K., Galasko D., et al., Reduction of β-amyloid peptide 42 in the cerebrospinal fluid of patients with Alzheimer’s disease, Ann. Neurol., 1995, 38, 643–648

  • [12] Blennow K., Hampel H., CSF markers for incipient Alzheimer’s disease, Lancet Neurol., 2003, 2, 605–613

  • [13] Sunderland T., Linker G., Mirza N., Putnam K.T., Friedman D.L., Kimmel L.H., et al., Decreased beta-amyloid1–42 and increased tau levels in cerebrospinal fluid of patients with Alzheimer disease, J. Am. Med. Assoc., 2003, 289, 2094–2103

  • [14] Verwey N.A., van der Flier W.M., Blennow K., Clark C., Sokolow S., De Deyn P.P., et al., A worldwide multicentre comparison of assays for cerebrospinal fluid biomarkers in Alzheimer’s disease, Ann. Clin. Biochem., 2009, 46, 235–240

  • [15] Hort J., Bratos A., Pirttila T., Scheltens P., Use of cerebrospinal fluid biomarkers in diagnosis of dementia across Europe, Eur. J. Neurol., 2010, 17, 90–96

  • [16] Mattsson N., Andreasson U., Persson S., Arai H., Batish S.D., Bernardini S., et al., The Alzheimer’s Association external quality control program for cerebrospinal fluid biomarkers, Alzheimers Dement., 2011, 7, 386–395

  • [17] Vanderstichele H., Bibl M., Engelborghs S., Le Bastard N., Lewczuk P., Molinuevo J.L., et al., Standardization of preanalytical aspects of cerebrospinal fluid biomarker testing for Alzheimer’s disease diagnosis: a consensus paper from the Alzheimer’s Biomarkers Standardization Initiative, Alzheimers Dement., 2012, 8, 65–73

  • [18] Teunissen C.E., Verwey N.A., Kester M.I., van Uffelen K., Blankenstein M.A., Standardization of assay procedures for analysis of the CSF biomarkers amyloid beta (1–42), tau, and phosphorylated tau in Alzheimer’s disease: report of an international workshop, Int. J. Alzheimers Dis., 2010, pii: 635053

  • [19] Boban M., Malojčić B., Mimica N., Vuković S., Zrilić I., Hof P.R., et al., The reliability and validity of the Mini-Mental State Examination in the elderly Croatian population, Dement. Geriatr. Cogn. Disord., 2012, 33, 385–392

  • [20] McKhann G., Drachman D., Folstein M., Katzman R., Price D., Stadlan E.M., Clinical diagnosis of Alzheimer’s disease: report of the NINCDSADRDA Work Group under the auspices of Department of Health and Human Services Task Force on Alzheimer’s Disease, Neurology, 1984, 34, 939–944

  • [21] Petersen R.C., Smith G.E., Waring S.C., Ivnik R.J., Tangalos E.G., Kokmen E., Mild cognitive impairment: clinical characterization and outcome, Arch. Neurol., 1999, 56, 303–308

  • [22] Bablok W., Passing H., Application of statistical procedures in analytical instrument testing, J. Automat. Chem., 1985, 7, 74–79

  • [23] Bilić-Zulle L., Comparison of methods: Passing and Bablok regression, Biochem. Med., 2011, 21, 49–52

  • [24] Mattsson N., Blennow H., Zetterberg H., Inter-laboratory variation in cerebrospinal fluid biomarkers for Alzheimer’s disease: united we stand, divided we fall, Clin. Chem. Lab. Med., 2010, 48, 603–607

  • [25] Verwey N.A., Bouwman F.H., van der Flier W.M., Veerhuis R., Scheltens P., Blankenstein M.A., Variability in longitudinal cerebrospinal fluid tau and phosphorylated tau measurements, Clin. Chem. Lab. Med., 2008, 46, 1300–1304

  • [26] Bjerke M., Portelius E., Minthon L., Wallin A., Anckarsäter H., Anckarsäter R., et al., Confounding factors influencing amyloid beta concentration in cerebrospinal fluid, Int. J. Alzheimers Dis., 2010, pii: 986310

  • [27] Teunissen C.E., Petzold A., Bennett J.L., Berven F.S., Brundin L., Comabella M., et al., A consensus protocol for the standardization of cerebrospinal fluid collection and biobanking, Neurology, 2009, 73, 1914–1922

  • [28] Fialova L., Bartos A., Svarcova J., Dolezil D., Malbohan I., Stanovení tau proteinu v mozkomíšním moku pacientů s roztroušenou sklerózou dvěma soupravami ELISA [Tau protein determination in cerebrospinal fluid in patients with multiple sclerosis by two ELISA kits], Klin. Biochem. Metab., 2011, 19, 113–118

  • [29] Regeniter A., Kuhle J., Baumann J., Sollberger M., Herdener M., Kunze U., et al., Biomarkers of dementia: comparison of electrochemiluminescence results and reference ranges with conventional ELISA, Methods, 2012, 56, 494–499

  • [30] Olsson A., Vanderstichele H., Andreasen N., De Meyer G., Wallin A., Holmberg B., et al., Simultaneous measurement of β-amyloid(1–42), total tau, and phosphorylated tau (Thr181) in cerebrospinal fluid by the xMAP technology, Clin. Chem., 2005, 51, 336–345

  • [31] Reijn T.S., Rikkert M.O., van Geel W.J., de Jong D., Verbeek M.M., Diagnostic accuracy of ELISA and xMAP technology for analysis of amyloid beta (42) and tau proteins, Clin. Chem., 2007, 53, 859–865

  • [32] Fagan A.M., Shaw L.M., Xiong C., Vanderstichele H., Mintun M.A., Trojanowski J.Q., et al., Comparison of analytical platforms for cerebrospinal fluid measures of Aβ1–42, total tau and p-tau181 for identifying Alzheimer’s disease amyloid plaque pathology, Arch. Neurol., 2011, 68, 1137–1144

  • [33] Wang L.-S., Leung Y.Y., Chang S.-K., Leight S., Knapik-Czajka M., Baek Y., et al., Comparison of xMAP and ELISA assays for detecting cerebrospinal fluid biomarkers of Alzheimer’s disease, J. Alzheimers Dis., 2012, 31, 439–445

  • [34] Le Bastard N., Coart E., Vanderstichele H., Vanmechelen E., Martin J.J., Engelborghs S., Comparison of two analytical platforms for the clinical qualification of Alzheimer’s disease biomarkers in pathologically-confirmed dementia, J. Alzheimers Dis., 2013, 33, 117–131

  • [35] Sjögren M., Vanderstichele H., Agren H., Zachrisson O., Edsbagge M., Wikkelsø C., et al., Tau and Abeta42 in cerebrospinal fluid from healthy adults 21–93 years of age: establishment of reference values, Clin. Chem., 2001, 47, 1776–1781

  • [36] Lewczuk P., Kornhuber J., Vanderstichele H., Vanmechelen E., Esselmann H., Bibl M., et al., Multiplexed quantification of dementia biomarkers in the CSF of patients with early dementias and MCI: a multicenter study, Neurobiol. Aging, 2008, 29, 812–818

  • [37] Shaw L.M., Vanderstichele H., Knapik-Czajka M., Figurski M., Coart E., Blennow K., et al., Qualification of the analytical and clinical performance of CSF biomarker analyses in ADNI, Acta Neuropathol., 2011, 121, 597–609

  • [38] Shaw L.M., Korecka M., Clark C.M., Lee V.M.-Y., Trojanowski J.Q., Biomarkers of neurodegeneration for diagnosis and monitoring therapeutics, Nat. Rev. Drug Discov., 2007, 6, 295–303


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