In Agglutination Inhibition Absence of Agglutination Is Diagnostic of Antigen

A modification of the agglutination reaction, called agglutination inhibition, provides a highly sensitive assay for small quantities of an antigen. For example, one of the early types of home pregnancy test kits included latex particles coated with human chorionic gonadotropin (HCG) and antibody to HCG (Figure 6-8). The addition of urine from a pregnant woman, which contained HCG, inhibited agglutination of the latex particles when the anti-HCG antibody was added; thus the absence of agglutination indicated pregnancy.

Agglutination inhibition assays can also be used to determine whether an individual is using certain types of illegal drugs, such as cocaine or heroin. A urine or blood sample is first incubated with antibody specific for the suspected drug. Then red blood cells (or other particles) coated with the drug are added. If the red blood cells are not agglutinated by the antibody, it indicates the sample contained an antigen recognized by the antibody, suggesting that the individual was

KIT REAGENTS

Hapten carrier-conjugate Anti-HCG antibody

TEST PROCEDURE

conjugate

Observe for visible clumping

POSSIBLE REACTIONS

reaction: not pregnant conjugate reaction: not pregnant

(+ reaction: pregnant

Agglutination Inhibition Hcg Assay

Visible clumping

No visible clumping

(+ reaction: pregnant

HCG in urine

Visible clumping

FIGURE 6-8

The original home pregnancy test kit employed hapten inhibition to determine the presence or absence of human chorionic gonadotropin (HCG). The original test kits used the presence or absence of visible clumping to determine whether HCG was present. If a woman was not pregnant, her urine would not contain HCG; in this case, the anti-HCG antibodies and HCG-carrier conjugate in the

No visible clumping kit would react, producing visible clumping. If a woman was pregnant, the HCG in her urine would bind to the anti-HCG antibodies, thus inhibiting the subsequent binding of the antibody to the HCG-carrier conjugate. Because of this inhibition, no visible clumping occurred if a woman was pregnant. The kits currently on the market use ELISA-based assays (see Figure 6-10).

using the illicit drug. One problem with these tests is that some legal drugs have chemical structures similar to those of illicit drugs, and these legal drugs may cross-react with the antibody, giving a false-positive reaction. For this reason a positive reaction must be confirmed by a nonimmunologic method.

Agglutination inhibition assays are widely used in clinical laboratories to determine whether an individual has been exposed to certain types of viruses that cause agglutination of red blood cells. If an individual's serum contains specific antiviral antibodies, then the antibodies will bind to the virus and interfere with hemagglutination by the virus. This technique is commonly used in premarital testing to determine the immune status of women with respect to rubella virus. The reciprocal of the last serum dilution to show inhibition of rubella hemagglutination is the titer of the serum. A titer greater than 10 (1:10 dilution) indicates that a woman is immune to rubella, whereas a titer of less than 10 is indicative of a lack of immunity and the need for immunization with the rubella vaccine.

Essentials of Human Physiology

Essentials of Human Physiology

This ebook provides an introductory explanation of the workings of the human body, with an effort to draw connections between the body systems and explain their interdependencies. A framework for the book is homeostasis and how the body maintains balance within each system. This is intended as a first introduction to physiology for a college-level course.

Get My Free Ebook


Responses

Post a comment