Chapter 6 : Human Immunodeficiency Virus

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Human Immunodeficiency Virus, Page 1 of 2

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This chapter focuses on the various technologies that are currently available from commercial companies for the diagnosis and monitoring of human immunodeficiency virus (HIV) infections, and it is not meant to duplicate more extensive reviews of HIV. Infection with HIV type 1 (HIV-1) results in the induction of a humoral antibody response specific to viral proteins, with the production of immunoglobulin A (IgA), IgM, and IgG. The majority of the automated immunoassay analyzers provide walk-away simplicity to perform assays from sample processing through interpretation of results. The major advantage of Western blot assays over enzyme immunoassays (EIAs) is that the specific interaction of antibody and antigen can be directly visualized. Immunofluorescence assay (IFA) is a very useful and inexpensive alternative to performing Western blot assays for confirmation of HIVspecific antibody responses. The Fluorognost HIV-1 IFA is based on the specific binding of HIV-1 antibodies in a specimen to HIV-1 antigens expressed on the surfaces of immortalized human T- cells fixed to glass slides. Specific antibody-antigen complexes are then detected using an anti-human antibody conjugated with fluorescein isothiocyanate. The development of molecular assays to quantitate the levels of HIV RNA in infected patients has provided one of the most valuable tools to assess the progression of HIV disease, monitor the impact of antiviral therapy, predict treatment failure and the emergence of drug-resistant viruses, and facilitate our understanding of the natural history and pathogenesis of this virus.

Citation: Hodinka R. 2002. Human Immunodeficiency Virus, p 100-127. In Truant A (ed), Manual of Commercial Methods in Clinical Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555817961.ch6

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Image of FIGURE 1

Schematic of a third-generation screening enzyme immunoassay for the detection of antibodies to HIV-1 and HIV-2. Abbreviations: rHIV, recombinant HIV; HRPO, horseradish peroxidase; OPD, -phenylenediamine.

Citation: Hodinka R. 2002. Human Immunodeficiency Virus, p 100-127. In Truant A (ed), Manual of Commercial Methods in Clinical Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555817961.ch6
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Image of FIGURE 2

Illustration of the steps involved in the Western blotting technique. Abbreviations: SDS, sodium dodecyl sulfate; PAGE, polyacrylamide gel electrophoresis; P, positive; N, negative; I, indeterminate.

Citation: Hodinka R. 2002. Human Immunodeficiency Virus, p 100-127. In Truant A (ed), Manual of Commercial Methods in Clinical Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555817961.ch6
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Image of FIGURE 3

Results of HIV-1-specific Western blot assays. Lane 1, high-positive (HP) control; lane 2, low-positive (LP) control; lane 3, negative (N) control; lanes 4 to 8, positive (P) sera; lane 9, indeterminate (I) serum. The numbers at the left refer to approximate molecular weights of HIV- 1 antigens. The criteria used for the interpretation of a positive HIV-1 Western blot is that of the Centers for Disease Control and Prevention and the Association of Public Health Laboratories and includes any two bands of p24, gp41, or gp160-gp120 ( ). For a specimen to be negative for HIV-1 antibodies by Western blotting, no banding pattern can be observed. Any banding pattern on a Western blot that does not meet the criteria of a positive result is interpreted as indeterminate for antibodies to HIV-1.

Citation: Hodinka R. 2002. Human Immunodeficiency Virus, p 100-127. In Truant A (ed), Manual of Commercial Methods in Clinical Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555817961.ch6
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Image of FIGURE 4

Schematic of an EIA for the detection of HIV-1 p24 antigen. Abbreviations: Ag, antigen; HRPO, horseradish peroxidase; OPD, -phenylenediamine.

Citation: Hodinka R. 2002. Human Immunodeficiency Virus, p 100-127. In Truant A (ed), Manual of Commercial Methods in Clinical Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555817961.ch6
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