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Chapter 9 : Serologic (Antibody Detection) Methods

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Serologic (Antibody Detection) Methods, Page 1 of 2

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Abstract:

For communicable diseases, clinical management, and public health response, it is often important to know the body's immune response following exposure and infection with pathogens. Although humoral and cell-mediated immunity both play roles in the body's specific immunity against viral pathogens, testing antibody response for humoral immunity is much more common and is also easier in clinical virology laboratories than testing for cell-mediated immunity because of the convenience of antibody serological testing methods. There is a long history of using various serologic methods in clinical virology laboratories for antibody detection. Some methods, such as complement fixation test and immunodiffusion test, have been gradually phased out and replaced by faster and less laborious methods (1, 2). In this chapter, we will focus on antibody detection methods used in clinical virology laboratories: neutralization, hemagglutination inhibition, indirect immunofluorescence, enzyme immunoassay, and Western blot.

Citation: Xia D, Wadford D, Preas C, Schnurr D. 2016. Serologic (Antibody Detection) Methods, p 105-116. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch9
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Figures

Image of FIGURE 1
FIGURE 1

Diagram of the functional aspects of a fluorescent microscope used to detect FITC-labeled antibodies. 1) Broad spectrum light from mercury arc lamp or fiber optic light source emits light of all wavelengths, 2) bandpass excitation filter allows ONLY 495 nm to pass through for FITC excitation, 3) dichroic mirror: reflects light at 495 nm; transmits light at all other wavelengths, 4) longpass emission filter (Barrier filter) allows >500 nm to pass through, and 5) eyepiece.

Citation: Xia D, Wadford D, Preas C, Schnurr D. 2016. Serologic (Antibody Detection) Methods, p 105-116. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch9
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Image of FIGURE 2
FIGURE 2

Sequential addition of reagents in the indirect EIA.

Citation: Xia D, Wadford D, Preas C, Schnurr D. 2016. Serologic (Antibody Detection) Methods, p 105-116. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch9
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Image of FIGURE 3
FIGURE 3

Sequential addition of reagents in the competitive EIA.

Citation: Xia D, Wadford D, Preas C, Schnurr D. 2016. Serologic (Antibody Detection) Methods, p 105-116. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch9
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Image of FIGURE 4
FIGURE 4

Sequential addition of reagents in the IgM capture EIA.

Citation: Xia D, Wadford D, Preas C, Schnurr D. 2016. Serologic (Antibody Detection) Methods, p 105-116. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch9
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Image of FIGURE 5
FIGURE 5

Depiction of rheumatoid factor (RF) false positive in indirect IgM EIA.

Citation: Xia D, Wadford D, Preas C, Schnurr D. 2016. Serologic (Antibody Detection) Methods, p 105-116. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch9
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Image of FIGURE 6
FIGURE 6

Depiction of heterophilic (Hp) antibody false positive in IgM capture EIA.

Citation: Xia D, Wadford D, Preas C, Schnurr D. 2016. Serologic (Antibody Detection) Methods, p 105-116. In Loeffelholz M, Hodinka R, Young S, Pinsky B (ed), Clinical Virology Manual, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819156.ch9
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