Laboratory Methods for the Diagnosis of Hereditary Amyloidoses

  • S. Michelle ShillerEmail author
  • Ahmet Dogan
  • Kimiyo M. Raymond
  • W. Edward HighsmithJr.
Part of the Current Clinical Pathology book series (CCPATH)


The laboratory assessment for hereditary amyloidosis is a multistep process beginning with the identification of amyloid in a biopsy specimen. Techniques to further confirm the presence of amyloid and direct subsequent confirmation of amyloid subtype include Congo Red staining, immunohistochemistry, laser capture tandem mass spectrometry, and gene sequencing. Relevant genes related to hereditary amyloidosis are also briefly summarized in this chapter.


Amyloid Mass spectrometry Gene sequencing Hereditary amyloidoses TTR amyloidosis Familial amyloidoses 



The authors would like to thank Steven R. Zeldenrust, MD, PhD Consultant, Division of Hematology, Mayo Clinic, Rochester, Minnesota, for his editorial help and clinical support; Jason Theis and Karen Schowalter for their assistance in the mass spectrometry and molecular genetics laboratories, respectively, at Mayo Clinic, Rochester, Minnesota; Tad Holtegaard and Cindy McFarlin for their technical support; and Debbie Johnson and Pam Carter for their administrative support.


  1. 1.
    Altland K, Benson MD, Costello CE, Ferlini A, Hazenberg BP, Hund E, et al. Genetic microheterogeneity of human transthyretin detected by IEF. Electrophoresis. 2007;28(12):2053–64.CrossRefPubMedGoogle Scholar
  2. 2.
    Connors LH, Lim A, Prokaeva T, Roskens VA, Costello CE. Tabulation of human transthyretin (TTR) variants, 2003. Amyloid. 2003;10(3):160–84.CrossRefPubMedGoogle Scholar
  3. 3.
    Maury CP, Baumann M. Isolation and characterization of cardiac amyloid in familial amyloid polyneuropathy type IV (Finnish): relation of the amyloid protein to variant gelsolin. Biochim Biophys Acta. 1990;1096(1):84–6.CrossRefPubMedGoogle Scholar
  4. 4.
    Lachmann HJ, Booth DR, Booth SE, Bybee A, Gilbertson JA, Gillmore JD, et al. Misdiagnosis of hereditary amyloidosis as AL (primary) amyloidosis. N Engl J Med. 2002;346(23):1786–91.CrossRefPubMedGoogle Scholar
  5. 5.
    Comenzo RL, Zhou P, Fleisher M, Clark B, Teruya-Feldstein J. Seeking confidence in the diagnosis of systemic AL (Ig light-chain) amyloidosis: patients can have both monoclonal gammopathies and hereditary amyloid proteins. Blood. 2006;107(9):3489–91.CrossRefPubMedGoogle Scholar
  6. 6.
    Bergen HR, Zeldenrust SR, Butz ML, Snow DS, Dyck PJ, Dyck PJ, et al. Identification of transthyretin variants by sequential proteomic and genomic analysis. Clin Chem. 2004;50(9):1544–52.CrossRefPubMedGoogle Scholar
  7. 7.
    Bergethon PR, Sabin TD, Lewis D, Simms RW, Cohen AS, Skinner M. Improvement in the polyneuropathy associated with familial amyloid polyneuropathy after liver transplantation. Neurology. 1996;47(4):944–51.CrossRefPubMedGoogle Scholar
  8. 8.
    Kishikawa M, Nakanishi T, Miyazaki A, Shimizu A, Nakazato M, Kangawa K, et al. Simple detection of abnormal serum transthyretin from patients with familial amyloidotic polyneuropathy by high-performance liquid chromatography/electrospray ionization mass spectrometry using material precipitated with specific antiserum. J Mass Spectrom. 1996;31(1):112–4.CrossRefPubMedGoogle Scholar
  9. 9.
    Theberge R, Connors L, Skare J, Skinner M, Falk RH, Costello CE. A new amyloidogenic transthyretin variant (Val122Ala) found in a compound heterozygous patient. Amyloid. 1999;6(1):54–8.CrossRefPubMedGoogle Scholar
  10. 10.
    Saraiva M. Transthyretin mutations in hyperthyroxinemia and amyloid diseases. Hum Mutat. 2001;17:493–503.CrossRefPubMedGoogle Scholar
  11. 11.
    Benson MD, Liepnieks J, Uemichi T, Wheeler G, Correa R. Hereditary renal amyloidosis associated with a mutant fibrinogen alpha-chain. Nat Genet. 1993;3(3):252–5.CrossRefPubMedGoogle Scholar
  12. 12.
    Cendron L, Trovato A, Seno F, Folli C, Alfieri B, Zanotti G, et al. Amyloidogenic potential of transthyretin variants: insights from structural and computational analyses. J Biol Chem. 2009;284(38):25832–41.PubMedCentralCrossRefPubMedGoogle Scholar
  13. 13.
    Jacob E, Edwards W, Zucker M, D’Cruz C, Seshan S, Crow F, et al. Homozygous transthyretin mutation in an African American Male. J Mol Diagn. 2007;9(1):127–31.PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Reddi HV, Jenkins S, Theis J, Thomas BC, Connors LH, Van Rhee F, et al. Homozygosity for the V122I mutation in transthyretin is associated with earlier onset of cardiac amyloidosis in the African American population in the seventh decade of life. J Mol Diagn. 2014;16(1):68–74.CrossRefPubMedGoogle Scholar
  15. 15.
    Eriksson M, Schonland S, Yumlu S, Hegenbart U, von Hutten H, Gioeva Z, et al. Hereditary apolipoprotein AI-associated amyloidosis in surgical pathology specimens: identification of three novel mutations in the APOA1 gene. J Mol Diagn. 2009;11(3):257–62.PubMedCentralCrossRefPubMedGoogle Scholar
  16. 16.
    Rowczenio D, Dogan A, Theis JD, Vrana JA, Lachmann HJ, Wechalekar AD, et al. Amyloidogenicity and clinical phenotype associated with five novel mutations in apolipoprotein A-I. Am J Pathol. 2011;179(4):1978–87. doi: 10.1016/j.ajpath.2011.06.024. Epub 2011 Aug 5.
  17. 17.
    Benson MD. The hereditary amyloidoses. Best Pract Res Clin Rheumatol. 2003;17(6):909–27.CrossRefPubMedGoogle Scholar
  18. 18.
    Kitagawa K, Wang J, Mastushita T, Kogishi K, Hosokawa M, Fu X, et al. Polymorphisms of mouse apolipoprotein A-II: seven alleles found among 41 inbred strains of mice. Amyloid. 2003;10(4):207–14.CrossRefPubMedGoogle Scholar
  19. 19.
    Lackner KJ, Law SW, Brewer Jr HB. The human apolipoprotein A-II gene: complete nucleic acid sequence and genomic organization. Nucleic Acids Res. 1985;13(12):4597–608.PubMedCentralCrossRefPubMedGoogle Scholar
  20. 20.
    Lee WM, Galbraith RM. The extracellular actin-scavenger system and actin toxicity. N Engl J Med. 1992;326(20):1335–41.CrossRefPubMedGoogle Scholar
  21. 21.
    de la Chapelle A, Tolvanen R, Boysen G, Santavy J, Bleeker-Wagemakers L, Maury CP, et al. Gelsolin-derived familial amyloidosis caused by asparagine or tyrosine substitution for aspartic acid at residue 187. Nat Genet. 1992;2(2):157–60.CrossRefPubMedGoogle Scholar
  22. 22.
    Meretoja J. Genetic aspects of familial amyloidosis with corneal lattice dystrophy and cranial neuropathy. Clin Genet. 1973;4(3):173–85.CrossRefPubMedGoogle Scholar
  23. 23.
    Paunio T, Kangas H, Kalkkinen N, Haltia M, Palo J, Peltonen L. Toward understanding the pathogenic mechanisms in gelsolin-related amyloidosis: in vitro expression reveals an abnormal gelsolin fragment. Hum Mol Genet. 1994;3(12):2223–9.CrossRefPubMedGoogle Scholar
  24. 24.
    Maury CP. Gelsolin-related amyloidosis. Identification of the amyloid protein in Finnish hereditary amyloidosis as a fragment of variant gelsolin. J Clin Investig. 1991;87(4):1195–9.PubMedCentralCrossRefPubMedGoogle Scholar
  25. 25.
    Sethi S, Theis JD, Quint P, Maierhofer W, Kurtin PJ, Dogan A, et al. Renal amyloidosis associated with a novel sequence variant of gelsolin. Am J Kidney Dis. 2013;61(1):161–6.CrossRefPubMedGoogle Scholar
  26. 26.
    Hamidi Asl L, Liepnieks JJ, Uemichi T, Rebibou JM, Justrabo E, Droz D, et al. Renal amyloidosis with a frame shift mutation in fibrinogen aalpha-chain gene producing a novel amyloid protein. Blood. 1997;90(12):4799–805.PubMedGoogle Scholar
  27. 27.
    Mourad G, Delabre JP, Garrigue V. Cardiac amyloidosis with the E526V mutation of the fibrinogen A alpha-chain. N Engl J Med. 2008;359(26):2847–8.CrossRefPubMedGoogle Scholar
  28. 28.
    Uemichi T, Liepnieks JJ, Yamada T, Gertz MA, Bang N, Benson MD. A frame shift mutation in the fibrinogen A alpha chain gene in a kindred with renal amyloidosis. Blood. 1996;87(10):4197–203.PubMedGoogle Scholar
  29. 29.
    Uemichi T, Liepnieks JJ, Benson MD. Hereditary renal amyloidosis with a novel variant fibrinogen. J Clin Invest. 1994;93(2):731–6.PubMedCentralCrossRefPubMedGoogle Scholar
  30. 30.
    Granel B, Serratrice J, Disdier P, Weiller PJ, Valleix S, Grateau G, et al. Underdiagnosed amyloidosis: amyloidosis of lysozyme variant. Am J Med. 2005;118(3):321–2.CrossRefPubMedGoogle Scholar
  31. 31.
    Granel B, Serratrice J, Valleix S, Grateau G, Droz D, Lafon J, et al. A family with gastrointestinal amyloidosis associated with variant lysozyme. Gastroenterology. 2002;123(4):1346–9.CrossRefPubMedGoogle Scholar
  32. 32.
    Yazaki M, Liepnieks JJ, Barats MS, Cohen AH, Benson MD. Hereditary systemic amyloidosis associated with a new apolipoprotein AII stop codon mutation Stop78Arg. Kidney Int. 2003;64(1):11–6.CrossRefPubMedGoogle Scholar
  33. 33.
    Benson MD. LECT2 amyloidosis. Kidney Int. 2010;77(9):757–9.CrossRefPubMedGoogle Scholar
  34. 34.
    Benson MD, James S, Scott K, Liepnieks JJ, Kluve-Beckerman B. Leukocyte chemotactic factor 2: a novel renal amyloid protein. Kidney Int. 2008;74(2):218–22.CrossRefPubMedGoogle Scholar
  35. 35.
    Murphy CL, Wang S, Kestler D, Larsen C, Benson D, Weiss DT, et al. Leukocyte chemotactic factor 2 (LECT2)-associated renal amyloidosis: a case series. Am J Kidney Dis. 2010;56(6):1100–7.PubMedCentralCrossRefPubMedGoogle Scholar
  36. 36.
    Yamagoe S, Kameoka Y, Hashimoto K, Mizuno S, Suzuki K. Molecular cloning, structural characterization, and chromosomal mapping of the human LECT2 gene. Genomics. 1998;48(3):324–9.CrossRefPubMedGoogle Scholar
  37. 37.
    Larsen CP, Walker PD, Weiss DT, Solomon A. Prevalence and morphology of leukocyte chemotactic factor 2-associated amyloid in renal biopsies. Kidney Int. 2010;77(9):816–9.CrossRefPubMedGoogle Scholar
  38. 38.
    Heegaard NH. beta(2)-microglobulin: from physiology to amyloidosis. Amyloid. 2009;16(3):151–73.CrossRefPubMedGoogle Scholar
  39. 39.
    Valleix S, Gillmore JD, Bridoux F, Mangione PP, Dogan A, Nedelec B, et al. Hereditary systemic amyloidosis due to Asp76Asn variant beta2-microglobulin. N Engl J Med. 2012;366(24):2276–83.CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • S. Michelle Shiller
    • 1
    • 2
    Email author
  • Ahmet Dogan
    • 3
  • Kimiyo M. Raymond
    • 1
  • W. Edward HighsmithJr.
    • 1
  1. 1.Department of Laboratory Medicine and PathologyMayo Clinic College of MedicineRochesterUSA
  2. 2.Department of Surgical PathologyBaylor University Medical CenterDallasUSA
  3. 3.Departments of Pathology and Laboratory MedicineMemorial Sloan Kettering Cancer CenterNew YorkUSA

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