1887

Chapter 87 : Antinuclear Antibody Tests

Authors: Alessandra Dellavance1, Wilson de Melo Cruvinel2, Paulo Luiz Carvalho Francescantonio3, Luis Eduardo Coelho Andrade4
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Affiliations: 1: Fleury Laboratories, Research and Development Department, Avenida Valdomiro de Lima 508, São Paulo, SP 04344-070, Brazil; 2: Pontifícia Universidade Católica de Goiás, School of Medical, Pharmaceutical and Biomedical Sciences, Avenida Universitária 1440, Setor Universitário, Goiânia, GO, 74.605-010, Brazil; 3: Pontifícia Universidade Católica de Goiás, School of Medical, Pharmaceutical and Biomedical Sciences, Avenida Universitária 1440, Setor Universitário, Goiânia, GO, 74.605-010, Brazil; 4: Escola Paulista de Medicina, Universidade Federal de Sao Paulo, Rheumatology Division, Rua Botucatu 740, Vila Clementino, Sao Paulo, SP 04023-062, and Fleury Laboratories, Immunology Division, Av. Valdomiro de Lima 508, São Paulo, SP 04344-070, Brazil
Content Type: Reference
Publication Year: 2016

Category: Immunology

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Abstract:

Autoantibodies are historical hallmarks in the establishment of the concept of autoimmunity and in the definition of the clinical limits of several autoimmune diseases. On a day-to-day basis, autoantibodies are helpful elements not only in the diagnosis of autoimmune diseases, but also frequently in the establishment of prognosis and in the monitoring of disease activity.

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
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Antinuclear Antibody Tests
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Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
/content/10.1128/9781555818722.ch87.ch87fig1
FIGURE 1

LE cell phenomenon. Viable neutrophils have their nucleus (thin arrow) displaced to the periphery of the cell due to the phagocytosis of a large piece of opsonized chromatin (wide arrow) from dead cells.

10.1128/9781555818722/8722ch87fig1_thmb.gif
10.1128/9781555818722/8722ch87fig1.gif
Image of FIGURE 1
FIGURE 1

LE cell phenomenon. Viable neutrophils have their nucleus (thin arrow) displaced to the periphery of the cell due to the phagocytosis of a large piece of opsonized chromatin (wide arrow) from dead cells.

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
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FIGURE 2

Influence of cell substrate on the ANA pattern. (A and B) Sample with anti-U1 RNP antibodies processed on slides from two different suppliers. The expected nuclear coarse speckled pattern is observed with the slide brand shown in panel A, whereas a nonspecific nuclear fine speckled pattern is observed the slide brand depicted in panel B. (C and D) Sample with anti-PCNA (proliferating cell nuclear antigen) antibody processed on slides from two different suppliers. The expected pleomorphic PCNA pattern is observed in the slide brand shown in panel C, whereas a nonspecific pattern is observed in the slide brand shown in panel D. Indirect immunofluorescence on HEp-2 cells; magnification, ×400.

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10.1128/9781555818722/8722ch87fig2.gif
Image of FIGURE 2
FIGURE 2

Influence of cell substrate on the ANA pattern. (A and B) Sample with anti-U1 RNP antibodies processed on slides from two different suppliers. The expected nuclear coarse speckled pattern is observed with the slide brand shown in panel A, whereas a nonspecific nuclear fine speckled pattern is observed the slide brand depicted in panel B. (C and D) Sample with anti-PCNA (proliferating cell nuclear antigen) antibody processed on slides from two different suppliers. The expected pleomorphic PCNA pattern is observed in the slide brand shown in panel C, whereas a nonspecific pattern is observed in the slide brand shown in panel D. Indirect immunofluorescence on HEp-2 cells; magnification, ×400.

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
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FIGURE 3

Checkerboard titration of the conjugate. A sample with known ANA titer is used to gauge the dilution of the conjugate that will reproduce the expected titer for that sample.

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10.1128/9781555818722/8722ch87fig3.gif
Image of FIGURE 3
FIGURE 3

Checkerboard titration of the conjugate. A sample with known ANA titer is used to gauge the dilution of the conjugate that will reproduce the expected titer for that sample.

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
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FIGURE 4

Example of report of the IIF-ANA test.

10.1128/9781555818722/8722ch87fig4_thmb.gif
10.1128/9781555818722/8722ch87fig4.gif
Image of FIGURE 4
FIGURE 4

Example of report of the IIF-ANA test.

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
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FIGURE 5

Panel of relevant IIF-ANA patterns. (A) Nuclear homogeneous pattern; (B) nuclear dense fine speckled pattern; (C) nuclear coarse speckled pattern; (D) nuclear SS-A/Ro pattern; (E) nuclear fine speckled pattern; (F) centromere pattern; (G) nuclear pleomorphic pattern; (H) ribosomal-P cytoplasmic pattern. Double arrows, cells in interphase; single arrow, cells in metaphase; arrowheads, cell nuclei with pleomorphic pattern and variable intensity. Indirect immunofluorescence using HEp-2 cells.

10.1128/9781555818722/8722ch87fig5_thmb.gif
10.1128/9781555818722/8722ch87fig5.gif
Image of FIGURE 5
FIGURE 5

Panel of relevant IIF-ANA patterns. (A) Nuclear homogeneous pattern; (B) nuclear dense fine speckled pattern; (C) nuclear coarse speckled pattern; (D) nuclear SS-A/Ro pattern; (E) nuclear fine speckled pattern; (F) centromere pattern; (G) nuclear pleomorphic pattern; (H) ribosomal-P cytoplasmic pattern. Double arrows, cells in interphase; single arrow, cells in metaphase; arrowheads, cell nuclei with pleomorphic pattern and variable intensity. Indirect immunofluorescence using HEp-2 cells.

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
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FIGURE 6

Pretest probability influences the diagnostic performance of tests. Anti-X antibody is a biomarker for the rare hypothetical disease Z and occurs in 1% of the general population. Indiscriminate testing for anti-X yields poor positive predictive value. Enrichment of the population sample to be tested according to clinical judgment restores the good performance of the anti-X antibody test.

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10.1128/9781555818722/8722ch87fig6.gif
Image of FIGURE 6
FIGURE 6

Pretest probability influences the diagnostic performance of tests. Anti-X antibody is a biomarker for the rare hypothetical disease Z and occurs in 1% of the general population. Indiscriminate testing for anti-X yields poor positive predictive value. Enrichment of the population sample to be tested according to clinical judgment restores the good performance of the anti-X antibody test.

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
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FIGURE 7

Decision-tree algorithm for classification of IIF-ANA patterns proposed by the IV Brazilian Consensus on ANA. MCP, metaphase chromatin plate.

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10.1128/9781555818722/8722ch87fig7.gif
Image of FIGURE 7
FIGURE 7

Decision-tree algorithm for classification of IIF-ANA patterns proposed by the IV Brazilian Consensus on ANA. MCP, metaphase chromatin plate.

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
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References

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Tables

Antinuclear Antibody Tests
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Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
/content/10.1128/9781555818722.ch87.ch87tab1
TABLE 1

Advantages of the ANA assay on HEp-2 cells

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TABLE 1

Advantages of the ANA assay on HEp-2 cells

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
/content/10.1128/9781555818722.ch87.ch87tab2
TABLE 2

Ability of the ANA-HEp-2 test to detect clinically relevant autoantibodies

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TABLE 2

Ability of the ANA-HEp-2 test to detect clinically relevant autoantibodies

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
/content/10.1128/9781555818722.ch87.ch87tab3
TABLE 3

Specific composite IIF-ANA patterns

Generic image for table
TABLE 3

Specific composite IIF-ANA patterns

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87
/content/10.1128/9781555818722.ch87.ch87tab4
TABLE 4

IIF-ANA patterns, targeted autoantigen, and disease associations

Generic image for table
TABLE 4

IIF-ANA patterns, targeted autoantigen, and disease associations

Citation: Dellavance A, Cruvinel W, Francescantonio P, Andrade L. 2016. Antinuclear Antibody Tests, p 843-858. 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.ch87

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