1887

Chapter 88 : Detection of Autoantibodies by Enzyme-Linked Immunosorbent Assay and Bead Assays

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Ebook: Choose a downloadable PDF or ePub file. Chapter is a downloadable PDF file. File must be downloaded within 48 hours of purchase

Buy this Chapter
Digital (?) $30.00

Preview this chapter:
Zoom in
Zoomout

Detection of Autoantibodies by Enzyme-Linked Immunosorbent Assay and Bead Assays, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555818722/9781555818715_CH88-1.gif /docserver/preview/fulltext/10.1128/9781555818722/9781555818715_CH88-2.gif

Abstract:

Autoantibodies directed against intracellular antigens are characteristic features of a number of human autoimmune diseases and certain malignancies (1–3). Studies of systemic autoimmune rheumatic diseases have provided strong evidence that autoantibodies are maintained by antigen-driven responses (4, 5) and that autoantibodies can be reporters from the immune system, revealing the identities of antigens involved in disease pathogenesis. Historically, autoantibody detection and analysis have relied on a number of different technologies, such as hemagglutination and particle aggregation, immunodiffusion, indirect immunofluorescence (IIF), complement fixation, counterimmunoelectrophoresis (CIE), Western and dot blotting, immunoprecipitation (IP), and enzyme-linked immunosorbent assay (ELISA), and on functional assays that demonstrate inhibition of the catalytic or other functional activity of the antigen of interest. These technologies have limitations because they tend to be labor-intensive and time-consuming, are limited in throughput, are semiquantitative, and are not adaptable to leading-edge research. Immunodiffusion has been used for over 50 years, and it is still used in some clinical laboratories because it is inexpensive and has high specificity, but it lacks sensitivity and can take up to 48 h before precipitin lines are interpretable. Western blotting is more costly and time-consuming, and not all autoantibodies are detected by this technique. For example, in the SS-A/Ro system, it has been observed that IP techniques are required to identify some sera that contain antibodies reacting only with the “native” SS-A/Ro particle (6). IP protocols that use extracts from [S]methionine-labeled cells are not suitable for the detection of all autoantibodies, such as antibodies to Ro52/TRIM21 protein (7). ELISA techniques have rapidly advanced, but highly specific, sensitive, and reliable assays that use highly purified or recombinant proteins are limited by intermanufacturer and interlaboratory variation of results (8). Immunodiffusion and CIE generally favor high-titer sera and often cannot discriminate multiple autoantibody responses that are characteristic of systemic autoimmune rheumatic disease sera.

Citation: Chan E, Burlingame R, Fritzler M. 2016. Detection of Autoantibodies by Enzyme-Linked Immunosorbent Assay and Bead Assays, p 859-867. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch88
Highlighted Text: Show | Hide
Loading full text...

Full text loading...

Figures

Image of FIGURE 1
FIGURE 1

Enhanced expression of recombinant proteins is illustrated by the effect on the bacterial host. A construct, pET-p115, was transformed into either JM109(DE3) or Rosetta(DE3), and recombinant proteins were produced with the transformed bacteria using the same culture medium (2YT broth) and conditions. To the cultured bacteria was added 1 mM isopropyl-b--thiogalactopyranoside at time zero, and cells were harvested as pellets after 1 or 2 h, solubilized in gel sample buffer, and analyzed with SDS-PAGE using a 5% gel. Proteins were detected with Coomassie brilliant blue G-250 stain. Note that the recombinant JM109 culture generated major products of ~60 kDa (arrowhead) plus other lower-molecular-mass products, whereas the Rosetta culture yielded major products (arrow) corresponding to the native protein. Lane M, molecular mass markers (numbers at the left are masses in kilodaltons).

Citation: Chan E, Burlingame R, Fritzler M. 2016. Detection of Autoantibodies by Enzyme-Linked Immunosorbent Assay and Bead Assays, p 859-867. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch88
Permissions and Reprints Request Permissions
Download as Powerpoint

References

/content/book/10.1128/9781555818722.ch88
1. Chan EKL, Andrade LEC. 1992. Antinuclear antibodies in Sjögren's syndrome. Rheum Dis Clin North Am 18:551570.[PubMed]
2. Satoh M, Vázquez-Del Mercado M, Chan EKL. 2009. Clinical interpretation of antinuclear antibody tests in systemic rheumatic diseases. Mod Rheumatol 19:219228.[CrossRef].[PubMed]
3. Mahler M, Pierangeli S, Meroni PL, Fritzler MJ. 2014. Autoantibodies in systemic autoimmune disorders. J Immunol Res 2014:263091.[CrossRef].[PubMed]
4. Tan EM, Chan EKL, Sullivan KF, Rubin RL. 1988. Antinuclear antibodies (ANAs): diagnostically specific immune markers and clues toward the understanding of systemic autoimmunity. Clin Immunol Immunopathol 47:121141.[PubMed].[CrossRef]
5. Radic MZ, Weigert M. 1994. Genetic and structural evidence for antigen selection of anti-DNA antibodies. Annu Rev Immunol 12:487520.[CrossRef].[PubMed]
6. Boire G, Lopez-Longo FJ, Lapointe S, Ménard HA. 1991. Sera from patients with autoimmune disease recognize conformational determinants on the 60-kd Ro/SS-A protein. Arthritis Rheum 34:722730.[PubMed].[CrossRef]
7. Chan EKL, Buyon JP,. 1994. The SS-A/Ro antigen, p 118. In van Venrooij WJ, Maini RN (ed), Manual of Biological Markers of Disease. Kluwer Academic Publishers, Dordrecht, Netherlands.
8. Tan EM, Smolen JS, McDougal JS, Fritzler MJ, Gordon T, Hardin JA, Kalden JR, Lahita RG, Maini RN, Reeves WH, Rothfield NF, Takasaki Y, Wiik A, Wilson M, Koziol JA. 2002. A critical evaluation of enzyme immunoassay kits for detection of antinuclear autoantibodies of defined specificities. II. Potential for quantitation of antibody content. J Rheumatol 29:6874.[PubMed]
9. Bizzaro N, Wiik A. 2004. Appropriateness in anti-nuclear antibody testing: from clinical request to strategic laboratory practice. Clin Exp Rheumatol 22:349355.[PubMed]
10. Wiik AS, Gordon TP, Kavanaugh AF, Lahita RG, Reeves W, van Venrooij WJ, Wilson MR, Fritzler M IUIS/WHO/AF/CDC Committee for the Standardization of Autoantibodies in Rheumatic and Related Diseases. 2004. Cutting edge diagnostics in rheumatology: the role of patients, clinicians, and laboratory scientists in optimizing the use of autoimmune serology. Arthritis Rheum 51:291298.[CrossRef].[PubMed]
11. Meroni PL, Schur PH. 2010. ANA screening: an old test with new recommendations. Ann Rheum Dis 69:14201422.[CrossRef].[PubMed]
12. Mahler M, Ngo JT, Schulte-Pelkum J, Luettich T, Fritzler MJ. 2008. Limited reliability of the indirect immunofluorescence technique for the detection of anti-Rib-P antibodies. Arthritis Res Ther 10:R131.[CrossRef].[PubMed]
13. Fritzler MJ, Wiik A, Fritzler ML, Barr SG. 2003. The use and abuse of commercial kits used to detect autoantibodies. Arthritis Res Ther 5:192201.[CrossRef].[PubMed]
14. Ou Y, Enarson P, Rattner JB, Barr SG, Fritzler MJ. 2004. The nuclear pore complex protein Tpr is a common autoantigen in sera that demonstrate nuclear envelope staining by indirect immunofluorescence. Clin Exp Immunol 136:379387.[CrossRef].[PubMed]
15. Chan EKL, Fritzler MJ (ed). 2013. Ten years of progress in GW/P body research. Springer, New York, NY.
16. Maecker HT, Lindstrom TM, Robinson WH, Utz PJ, Hale M, Boyd SD, Shen-Orr SS, Fathman CG. 2012. New tools for classification and monitoring of autoimmune diseases. Nat Rev Rheumatol 8:317328.[CrossRef].[PubMed]
17. Mahler M, Meroni PL, Bossuyt X, Fritzler MJ. 2014. Current concepts and future directions for the assessment of autoantibodies to cellular antigens referred to as anti-nuclear antibodies. J Immunol Res 2014:315179.[CrossRef].[PubMed]
18. Chan-Hui PY, Stephens K, Warnock RA, Singh S. 2004. Applications of eTag trade mark assay platform to systems biology approaches in molecular oncology and toxicology studies. Clin Immunol 111:162174.[CrossRef].[PubMed]
19. Illei GG, Tackey E, Lapteva L, Lipsky PE. 2004. Biomarkers in systemic lupus erythematosus. I. General overview of biomarkers and their applicability. Arthritis Rheum 50:17091720.[CrossRef].[PubMed]
20. Kantor AB, Wang W, Lin H, Govindarajan H, Anderle M, Perrone A, Becker C. 2004. Biomarker discovery by comprehensive phenotyping for autoimmune diseases. Clin Immunol 111:186195.[CrossRef].[PubMed]
21. Sharp V, Utz PJ. 2007. Technology insight: can autoantibody profiling improve clinical practice? Nat Clin Pract Rheumatol 3:96103.[CrossRef].[PubMed]
22. Robinson WH, DiGennaro C, Hueber W, Haab BB, Kamachi M, Dean EJ, Fournel S, Fong D, Genovese MC, de Vegvar HE, Skriner K, Hirschberg DL, Morris RI, Muller S, Pruijn GJ, van Venrooij WJ, Smolen JS, Brown PO, Steinman L, Utz PJ. 2002. Autoantigen microarrays for multiplex characterization of autoantibody responses. Nat Med 8:295301.[CrossRef].[PubMed]
23. Thiel A, Scheffold A, Radbruch A. 2004. Antigen-specific cytometry—new tools arrived! Clin Immunol 111:155161.[CrossRef].[PubMed]
24. Mahler M, Radice A, Yang W, Bentow C, Seaman A, Bianchi L, Sinico RA. 2012. Development and performance evaluation of novel chemiluminescence assays for detection of anti-PR3 and anti-MPO antibodies. Clin Chim Acta 413:719726.[CrossRef].[PubMed]
25. Fritzler MJ, Conrad K, Meurer M, Sack U, Shoenfeld Y,. 2002. New technologies in the detection of autoantibodies, p 5063. In Conrad K (ed), Autoantigens, Autoantibodies, Autoimmunity. Pabst Scientific Publishers, Lengerich, Germany.
26. Joos TO, Stoll D, Templin MF. 2002. Miniaturised multiplexed immunoassays. Curr Opin Chem Biol 6:7680.[PubMed].[CrossRef]
27. Xue Q, Wainright A, Gangakhedkar S, Gibbons I. 2001. Multiplexed enzyme assays in capillary electrophoretic single-use microfluidic devices. Electrophoresis 22:40004007.[CrossRef].[PubMed]
28. Templin MF, Stoll D, Bachmann J, Joos TO. 2004. Protein microarrays and multiplexed sandwich immunoassays: what beats the beads? Comb Chem High Throughput Screen 7:223229.[PubMed].[CrossRef]
29. Utz PJ. 2004. “Hot technologies” for clinical immunology research. Clin Immunol 111:153154.[CrossRef].[PubMed]
30. Olsen NJ, Li QZ, Quan J, Wang L, Mutwally A, Karp DR. 2012. Autoantibody profiling to follow evolution of lupus syndromes. Arthritis Res Ther 14:R174.[CrossRef].[PubMed]
31. Thibault DL, Chu AD, Graham KL, Balboni I, Lee LY, Kohlmoos C, Landrigan A, Higgins JP, Tibshirani R, Utz PJ. 2008. IRF9 and STAT1 are required for IgG autoantibody production and B cell expression of TLR7 in mice. J Clin Invest 118:14171426.[CrossRef].[PubMed]
32. Ou Y, Sun D, Sharp GC, Hoch SO. 1997. Screening of SLE sera using purified recombinant Sm-D1 protein from a baculovirus expression system. Clin Immunol Immunopathol 83:310317.[PubMed].[CrossRef]
33. Schulte-Pelkum J, Fritzler M, Mahler M. 2009. Latest update on the Ro/SS-A autoantibody system. Autoimmun Rev 8:632637.[CrossRef].[PubMed]
34. Satoh M, Richards HB, Hamilton KJ, Reeves WH. 1997. Human anti-nuclear ribonucleoprotein antigen autoimmune sera contain a novel subset of autoantibodies that stabilizes the molecular interaction of U1RNP-C protein with the Sm core proteins. J Immunol 158:50175025.[PubMed]
35. Burlingame RW, Boey ML, Starkebaum G, Rubin RL. 1994. The central role of chromatin in autoimmune responses to histones and DNA in systemic lupus erythematosus. J Clin Invest 94:184192.[CrossRef].[PubMed]
36. Burlingame RW, Rubin RL. 1990. Subnucleosome structures as substrates in enzyme-linked immunosorbent assays. J Immunol Methods 134:187199.[PubMed].[CrossRef]
37. Kessenbrock K, Raijmakers R, Fritzler MJ, Mahler M. 2007. Synthetic peptides: the future of patient management in systemic rheumatic diseases? Curr Med Chem 14:28312838.[PubMed].[CrossRef]
38. Elkon K, Skelly S, Parnassa A, Moller W, Danho W, Weissbach H, Brot N. 1986. Identification and chemical synthesis of a ribosomal protein antigenic determinant in systemic lupus erythematosus. Proc Natl Acad Sci U S A 83:74197423.[PubMed].[CrossRef]
39. Mahler M, Stinton LM, Fritzler MJ. 2005. Improved serological differentiation between systemic lupus erythematosus and mixed connective tissue disease by use of an SmD3 peptide-based immunoassay. Clin Diagn Lab Immunol 12:107113.[CrossRef].[PubMed]
40. Mahler M. 2011. Sm peptides in differentiation of autoimmune diseases. Adv Clin Chem 54:109128.[PubMed].[CrossRef]
41. Schellekens GA, Visser H, de Jong BA, van den Hoogen FH, Hazes JM, Breedveld FC, van Venrooij WJ. 2000. The diagnostic properties of rheumatoid arthritis antibodies recognizing a cyclic citrullinated peptide. Arthritis Rheum 43:155163.[CrossRef].[PubMed]
42. Kato T, Miyakawa H, Ishibashi M. 2004. Frequency and significance of anti-glutathione S-transferase autoantibody (anti-GST A1-1) in autoimmune hepatitis. J Autoimmun 22:211216.[CrossRef].[PubMed]
43. Nozawa K, Fritzler MJ, von Mühlen CA, Chan EKL. 2004. Giantin is the major Golgi autoantigen in human anti-Golgi complex sera. Arthritis Res Ther 6:R95R102.[CrossRef].[PubMed]
44. Qiagen . 2003. The QIAexpressionist. A Handbook for High-Level Expression and Purification of 6×His-Tagged Proteins. Qiagen, Hilden, Germany.
45. Zhang JY, Casiano CA, Peng XX, Koziol JA, Chan EKL, Tan EM. 2003. Enhancement of antibody detection in cancer using panel of recombinant tumor-associated antigens. Cancer Epidemiol Biomarkers Prev 12:136143.[PubMed]
46. Fritzler MJ, Miller BJ. 1995. Detection of autoantibodies to SS-A/Ro by indirect immunofluorescence using a transfected and overexpressed human 60 kD Ro autoantigen in HEp-2 cells. J Clin Lab Anal 9:218224.[PubMed].[CrossRef]
47. Keech CL, McCluskey J, Gordon TP. 1994. Transfection and overexpression of the human 60-kDa Ro/SS-A autoantigen in HEp-2 cells. Clin Immunol Immunopathol 73:146151.[PubMed].[CrossRef]
48. Melegari A, Bonaguri C, Russo A, Luisita B, Trenti T, Lippi G. 2012. A comparative study on the reliability of an automated system for the evaluation of cell-based indirect immunofluorescence. Autoimmun Rev 11:713716.[CrossRef].[PubMed]
49. Meheus L, van Venrooij WJ, Wiik A, Charles PJ, Tzioufas AG, Meyer O, Steiner G, Gianola D, Bombardieri S, Union A, De Keyser S, Veys E, De Keyser F. 1999. Multicenter validation of recombinant, natural and synthetic antigens used in a single multiparameter assay for the detection of specific anti-nuclear autoantibodies in connective tissue disorders. Clin Exp Rheumatol 17:205214.[PubMed]
50. Yang JJ, Kim MH, Lee WI, Kang SY. 2014. Relevance of indirect immunofluorescence patterns and autoantibodies identified via line immunoassay in patients with rheumatoid arthritis. Lab Med 45:2531.[PubMed].[CrossRef]
51. Scussel-Lonzetti L, Joyal F, Raynauld JP, Roussin A, Rich E, Goulet JR, Raymond Y, Senécal JL. 2002. Predicting mortality in systemic sclerosis: analysis of a cohort of 309 French Canadian patients with emphasis on features at diagnosis as predictive factors for survival. Medicine (Baltimore) 81:154167.[PubMed].[CrossRef]
52. Radzimski C, Probst C, Teegen B, Rentzsch K, Blöcker IM, Dähnrich C, Schlumberger W, Stöcker W, Bogdanos DP, Komorowski L. 2013. Development of a recombinant cell-based indirect immunofluorescence assay for the determination of autoantibodies against soluble liver antigen in autoimmune hepatitis. Clin Dev Immunol 2013:572815.[CrossRef].[PubMed]
53. Fritzler MJ. 2012. Toward a new autoantibody diagnostic orthodoxy: understanding the bad, good and indifferent. Auto Immun Highlights 3:5158.[CrossRef].[PubMed]
54. Earley MC, Vogt RF Jr, Shapiro HM, Mandy FF, Kellar KL, Bellisario R, Pass KA, Marti GE, Stewart CC, Hannon WH. 2002. Report from a workshop on multianalyte microsphere assays. Cytometry 50:239242.[CrossRef].[PubMed]
55. Gilburd B, Abu-Shakra M, Shoenfeld Y, Giordano A, Bocci EB, delle Monache F, Gerli R. 2004. Autoantibodies profile in the sera of patients with Sjogren's syndrome: the ANA evaluation—a homogeneous, multiplexed system. Clin Dev Immunol 11:5356.[PubMed].[CrossRef]
56. Rouquette AM, Desgruelles C, Laroche P. 2003. Evaluation of the new multiplexed immunoassay, FIDIS, for simultaneous quantitative determination of antinuclear antibodies and comparison with conventional methods. Am J Clin Pathol 120:676681.[CrossRef].[PubMed]
57. Seideman J, Peritt D. 2002. A novel monoclonal antibody screening method using the Luminex-100 microsphere system. J Immunol Methods 267:165171.[PubMed].[CrossRef]
58. Jia XC, Raya R, Zhang L, Foord O, Walker WL, Gallo ML, Haak-Frendscho M, Green LL, Davis CG. 2004. A novel method of multiplexed competitive antibody binning for the characterization of monoclonal antibodies. J Immunol Methods 288:9198.[CrossRef].[PubMed]
59. Lynch HE, Sanchez AM, D'Souza MP, Rountree W, Denny TN, Kalos M, Sempowski GD. 2014. Development and implementation of a proficiency testing program for Luminex bead-based cytokine assays. J Immunol Methods 409:6271.[CrossRef].[PubMed]
60. Selak S, Fritzler MJ. 2004. Altered neurological function in mice immunized with early endosome antigen 1. BMC Neurosci 5:2.[CrossRef].[PubMed]
61. Selak S, Mahler M, Miyachi K, Fritzler ML, Fritzler MJ. 2003. Identification of the B-cell epitopes of the early endosome antigen 1 (EEA1). Clin Immunol 109:154164.[PubMed].[CrossRef]
62. Eystathioy T, Chan EKL, Mahler M, Luft LM, Fritzler ML, Fritzler MJ. 2003. A panel of monoclonal antibodies to cytoplasmic GW bodies and the mRNA binding protein GW182. Hybrid Hybridomics 22:7986.[CrossRef].[PubMed]
63. Eystathioy T, Chan EK, Takeuchi K, Mahler M, Luft LM, Zochodne DW, Fritzler MJ. 2003. Clinical and serological associations of autoantibodies to GW bodies and a novel cytoplasmic autoantigen GW182. J Mol Med (Berl) 81:811818.[CrossRef].[PubMed]
64. Miyachi K, Hirano Y, Horigome T, Mimori T, Miyakawa H, Onozuka Y, Shibata M, Hirakata M, Suwa A, Hosaka H, Matsushima S, Komatsu T, Matsushima H, Hankins RW, Fritzler MJ. 2004. Autoantibodies from primary biliary cirrhosis patients with anti-p95c antibodies bind to recombinant p97/VCP and inhibit in vitro nuclear envelope assembly. Clin Exp Immunol 136:568573.[CrossRef].[PubMed]
65. Fritzler MJ, Hanson C, Miller J, Eystathioy T. 2002. Specificity of autoantibodies to SS-A/Ro on a transfected and overexpressed human 60 kDa Ro autoantigen substrate. J Clin Lab Anal 16:103108.[PubMed].[CrossRef]
66. Mahler M, Kessenbrock K, Raats J, Fritzler MJ. 2004. Technical and clinical evaluation of anti-ribosomal P protein immunoassays. J Clin Lab Anal 18:215223.[CrossRef].[PubMed]
67. Mahler M, Kessenbrock K, Raats J, Williams R, Fritzler MJ, Blüthner M. 2003. Characterization of the human autoimmune response to the major C-terminal epitope of the ribosomal P proteins. J Mol Med (Berl) 81:194204.[CrossRef].[PubMed]
68. Mahler M, Fritzler MJ, Blüthner M. 2005. Identification of a SmD3 epitope with a single symmetrical dimethylation of an arginine residue as a specific target of a subpopulation of anti-Sm antibodies. Arthritis Res Ther 7:R19R29.[CrossRef].[PubMed]
69. Gussin HA, Ignat GP, Varga J, Teodorescu M. 2001. Anti-topoisomerase I (anti-Scl-70) antibodies in patients with systemic lupus erythematosus. Arthritis Rheum 44:376383.[CrossRef].[PubMed]
70. Venables PJ. 1997. Antibodies to Jo-1 and Ro-52: why do they go together? Clin Exp Immunol 109:403405.[CrossRef]
71. Bentow C, Swart A, Wu J, Seaman A, Manfredi M, Infantino M, Benucci M, Lakos G, Mahler M. 2013. Clinical performance evaluation of a novel rapid response chemiluminescent immunoassay for the detection of autoantibodies to extractable nuclear antigens. Clin Chim Acta 424:141147.[CrossRef].[PubMed]
72. Mahler M, Radice A, Sinico RA, Damoiseaux J, Seaman A, Buckmelter K, Vizjak A, Buchner C, Binder WL, Fritzler MJ, Cui Z. 2012. Performance evaluation of a novel chemiluminescence assay for detection of anti-GBM antibodies: an international multicenter study. Nephrol Dial Transplant 27:243252.[CrossRef].[PubMed]

Tables

Generic image for table
TABLE 1

Comparison of results of ELISA and ALBIA detection of anti-ribosomal P autoantibodies in 30 sera

Citation: Chan E, Burlingame R, Fritzler M. 2016. Detection of Autoantibodies by Enzyme-Linked Immunosorbent Assay and Bead Assays, p 859-867. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch88
Generic image for table
TABLE 2

Frequency of autoantibodies detected by the INOVA Quanta Plex assay in cohorts of patients with systemic rheumatic diseases and multiple sclerosis and in controls

Citation: Chan E, Burlingame R, Fritzler M. 2016. Detection of Autoantibodies by Enzyme-Linked Immunosorbent Assay and Bead Assays, p 859-867. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch88
Generic image for table
TABLE 3

Advantages, challenges, and opportunities relating to new array technologies

Citation: Chan E, Burlingame R, Fritzler M. 2016. Detection of Autoantibodies by Enzyme-Linked Immunosorbent Assay and Bead Assays, p 859-867. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch88

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error