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Chapter 94 : Principles and Procedures of Human Immunodeficiency Virus Serodiagnosis

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Abstract:

An important early impetus for the development and testing of serologic methods to detect human immunodeficiency virus (HIV) infection was the need to guarantee the safety of the blood supply. A quick review of the statistics provided in this chapter makes clear the need for the continued development of serologic methods, including point-of-care testing, that are useful especially in resource-poor settings. The standard screening tests for HIV infection involve detection of HIV-specific antibodies. The enzyme-linked immunosorbent assay (ELISA) is the standard screening test for HIV-1 infection. Individuals with indeterminate or positive ELISA results should undergo confirmation testing (generally by Western blotting [immunoblotting]) to determine if the reactivity is primarily directed to HIV-1 infection or secondarily directed to cross-reacting antibodies. It has now become the standard practice in blood banks to screen donated blood samples with one of the new HIV-1-HIV-2 combination ELISAs. The major core protein of HIV-1 is p24. The assay to detect the p24 antigen has been used for many years in the prognostic staging and management of infected individuals and is licensed in the United States as part of routine blood donor screening. Serologic assays to detect antibodies to HIV are the most widely used means of laboratory diagnosis of HIV infection. Most HIV testing algorithms include an ELISA as the screening component and Western blotting as the confirmatory method.

Citation: Dewar R, Highbarger H, Davey R, Metcalf J. 2006. Principles and Procedures of Human Immunodeficiency Virus Serodiagnosis, p 834-846. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch94

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Figures

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FIGURE 1

Diagrams of HIV-I virion structure (A) and the HIV-1 viral genome (B).

Citation: Dewar R, Highbarger H, Davey R, Metcalf J. 2006. Principles and Procedures of Human Immunodeficiency Virus Serodiagnosis, p 834-846. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch94
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Image of FIGURE 2
FIGURE 2

Hypothetical time course for the appearance of p24 antigen and HIV-1 antibodies (Ab) in serum during HIV-1 infection.

Citation: Dewar R, Highbarger H, Davey R, Metcalf J. 2006. Principles and Procedures of Human Immunodeficiency Virus Serodiagnosis, p 834-846. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch94
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Image of FIGURE 3
FIGURE 3

Algorithm for the correct use and sequence of serologic testing in the diagnosis of HIV infection.

Citation: Dewar R, Highbarger H, Davey R, Metcalf J. 2006. Principles and Procedures of Human Immunodeficiency Virus Serodiagnosis, p 834-846. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch94
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Image of FIGURE 4
FIGURE 4

Illustration of various patterns of positive HIV-1 Western blots, showing antibody banding at the major viral proteins that can be identified using this technique. For comparison, a negative Western blot and two examples of indeterminate Western blots are also shown.

Citation: Dewar R, Highbarger H, Davey R, Metcalf J. 2006. Principles and Procedures of Human Immunodeficiency Virus Serodiagnosis, p 834-846. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch94
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Image of FIGURE 5
FIGURE 5

The principle of the IFA test. FITC, fluorescein isothiocyanate; Ab, antibody.

Citation: Dewar R, Highbarger H, Davey R, Metcalf J. 2006. Principles and Procedures of Human Immunodeficiency Virus Serodiagnosis, p 834-846. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch94
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Tables

Generic image for table
TABLE 1

Commercial kits for HIV testing

Citation: Dewar R, Highbarger H, Davey R, Metcalf J. 2006. Principles and Procedures of Human Immunodeficiency Virus Serodiagnosis, p 834-846. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch94

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