LUSAKA APEX MEDICAL UNIVERSITY

FACULTY OF NURSING AND MIDWIFERY SCIENCES (FNMS)

DIPLOMA NURSING JANUARY 2016 INTAKE

SEROLOGICAL TESTING

Dr Simpokolwe K.
 INTRODUCTION

• Serology as a science began in 1901. Austrian

American immunologist Karl Landsteiner
(1868-1943) identified groups of red blood
cells as A, B, and O.
• Hence from the discovery ,cells of all types(e.g
blood cells, body cells and microorganisms)
carry proteins and other molecules on their
surface that are recognized by cells of the
immune system.
 SEROLOGY
• Serology: The study of blood serum,
with emphasis on testing of antibodies
in the serum.

• Antigen: A substance which stimulates

the body to produce an antibody.
Certain antigens found in the plasma
of the red blood cell’s membrane
account for blood type.

• Antibody: A protein molecule

produced by the body’s immune
system in response to a specific
antigen.

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 SEROLOGICAL TARGETS

Serology methods utilise the reactions and properties of serum

• ANTIBODIES
(Use of commercially available antigens)

• ANTIGENS
(Use of specific antibodies)
 DEFINITION OF TERMS

• Normally sterile site: sites in the human body that are normally free
from organisms or foreign material, e.g. blood, joint, brain.
• Unsterile site: sites in the human body that generally harbor
microorganisms, e.g. gut, oral cavity, nose, skin
• Specimen: a sample of tissue (e.g blood, urine) that may or may not
contain organisms.
• Isolate: a population of organisms (bacteria) that has been separated
from a mixture.
• Serotype: a group of closely related organisms with distinct
characteristics.

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 WHY USE SEROLOGY

1. Antigens and antibodies are easier to detect than

finding the organism directly.

2. Antigens and antibodies are produced in large

quantities and can be found in body fluids (e.g
blood, CSF, urine)

3. Culture is often problematic, time consuming and

insensitive due to the low concentration of the
organism in tissue.
 AVAILABLE TESTS
Antibodies
• Immunodiffusion
• Radioallergosorbent Test (RAST)
Antigens
• Latex Agglutination

• Radioimmunoassay (RIA)
Antibodies and antigens
• Enzyme-linked immunosorbent assay (ELISA)
• Enzyme Immunoassay (EIA)
BLOOD TYPING

Blood type A has antigen A on the surface of the cell and will
agglutinate with blood type B.
Blood type B has antigen B on the surface of the cell and will
agglutinate with blood type A.
Blood type AB has antigens A and B on the surface of the cells and
will not agglutinate with either type A or type B blood.
Blood type O has neither antigen A nor B and will not agglutinate.
BLOOD GROUPS

Type Antigen Antibody Can Give Can Get

Blood To Blood From

A A B A, AB O, A

B B A B, AB O, B

Neither
AB A and B AB A, B, O, AB
A nor B

O Neither A and B A, B, O, AB O
A nor B
BLOOD CHARACTERISTICS
Plasma is the fluid portion of the blood (55
percent).

Cells (45 percent)

Erythrocytes are red blood cells.

Leukocytes are the white blood cells

Thrombocytes or platelets are responsible
for blood clotting.
Serum is the liquid that separates from the
blood when a clot is formed.
 ANTIBODIES
• IgM: First antibody to appear in response to a
foreign substance exposure.

• IgG: The only antibody that crosses the

placenta to provide immunity to the fetus.

• Titer: The amount of antibodies present in the

blood, usually as a result of infection.

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 ANTIBODY TESTING

Advantages
1. Screening tool.
2. Readily available.
3. Indicates response to antigen (even if antigen
is not detectable) – may indicate infection or
immunity.

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Disadvantages
1. Paired testing necessary for some diseases.
2. Can’t differentiate between immunity and
disease.
3. Sensitivity and specificity:

False-negative result: compromised immune
system

False-positive result: liver disease, low disease
prevalence
 SEROLOGICAL TESTS
1. PRECIPITATION TESTS

2. AGGLUTINATION TESTS

3. NEUTRALISATION TESTS

4. ANTIGEN/AB CONJUGATION

5. ELISA TESTS

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PRECIPITATION TESTS

• One of the properties of some antibody

classes is the ability to precipitate from
solution when combined with multivalent
antigens; such reactions can be visualized.

• This behavior is called precipitation

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AGGLUTINATION TESTS

• Antibodies can agglutinate multivalent

particulate antigens, such as red blood cells or
bacteria
• This behavior is called agglutination.
• Serological tests based on agglutination are
usually more sensitive than those based on
precipitation.

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NEUTRALISATION TESTS

• Usually used for viral or bacterial/toxin

identification.
• Antibody neutralises the toxin and prevents
its action (antitoxins).
• Requires an indicator system e.g. lab animals,
tissue culture, etc

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ANTIGEN/AB CONJUGATION

• Antigens (or immunoglobulins) can be

conjugated with other molecules
(radioisotopes, enzymes or fluorescent dyes)
so that antigen-antibody binding can be
detected at extremely low concentrations.
• Examples are radioimmunoassay or enzyme-
immunoassay, Fluorescent antibody tests
(FAT).

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ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)

• Enzyme-Linked ImmunoSorbent Assay is a biochemical

technique used to detect the presence of an antibody or an
antigen in a sample of serum.

• In ELISA an unknown amount of antigen is affixed to a surface,

and then a specific antibody is washed over the surface so
that it can bind the antigen. This antibody is linked to an
enzyme, and in the final step a substance is added that the
enzyme can convert to some detectable signal.

• In fluorescence ELISA, when light is shone upon the sample,

any antigen/antibody complexes will fluoresce so that the
amount of antigen in the sample can be measured.
ELISA
Intensity of color correspond to
concentration of antibody.
 APPLICATIONS OF SEROLOGY

1. HIV testing.

2. Serum HCG (pregnancy).

3. Tests for hepatitis antigens and antibodies

4. Antibodies to bacteria.

5. Forensics