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Questions and Answers
What is the primary purpose of counter immunoelectrophoresis (CIE)?
What is the primary purpose of counter immunoelectrophoresis (CIE)?
To detect the binding of an antibody to its specific antigen.
Describe the role of the electric field in the CIE technique.
Describe the role of the electric field in the CIE technique.
The electric field accelerates the migration of antigens towards the anode and antibodies towards the cathode.
What does the presence of a precipitin line in CIE indicate?
What does the presence of a precipitin line in CIE indicate?
It indicates a positive reaction or a specific antigen-antibody interaction.
List one advantage of using CIE over double immunodiffusion.
List one advantage of using CIE over double immunodiffusion.
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What limitations are associated with the CIE technique?
What limitations are associated with the CIE technique?
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Mention two types of fluids in which CIE can be useful for diagnosis.
Mention two types of fluids in which CIE can be useful for diagnosis.
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What does the absence of a precipitin line suggest in CIE?
What does the absence of a precipitin line suggest in CIE?
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Explain the significance of multiple precipitin lines observed in a CIE test.
Explain the significance of multiple precipitin lines observed in a CIE test.
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Study Notes
Counter Immunoelectrophoresis (CIE)
- CIE is a laboratory technique used to detect the binding of an antibody to its specific antigen.
- An electric field is applied to accelerate the migration of antigens and antibodies in a gel medium (typically agar).
- The rapid movement of antibody and antigen causes them to meet and form a precipitation line.
Principle of CIE
- CIE relies on the movement of charged molecules. Strongly negatively charged antigens move towards the anode, while antibodies move towards the cathode.
- The test is performed on a glass slide with agarose gel. Two wells are punched out. One well holds the antigen, the other holds the antibody.
- An electric current is applied causing rapid migration.
- A visible precipitation line (precipitin arcs or lines) forms within 30-60 minutes, indicating a positive reaction.
Interpretation of Results
- A precipitin line between the antigen and antisera wells signifies a positive reaction, indicating a specific antigen-antibody reaction. (Specificity)
- The absence of these lines may indicate no reaction or the lack of a particular antibody for the antigen. (Non-specificity)
- Multiple precipitin lines suggest heterogeneity of the antibody reacting with the antigen.
Advantages and Limitations of CIE
- Advantages: Faster and more sensitive than double immunodiffusion (takes ~30 minutes).
- Limitations: More expensive than agglutination-based tests. Potentially lower sensitivity, speed, and simplicity compared to latex agglutination. Requires a larger quantity of antigen and antibody.
Uses of CIE
- Detects antigens and antibodies in serum, cerebrospinal fluid, and other bodily fluids to diagnose various infectious diseases (e.g., bacterial, viral, fungal, parasitic).
- Used for detecting Hepatitis B surface antigen (HBsAg), fetoprotein, hydatid, and amoebic antigens.
- Used for detecting pneumococcal capsular antigens in sputum.
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Description
Explore the fundamentals of Counter Immunoelectrophoresis (CIE), a laboratory technique that detects antibody-antigen binding through electric field-induced migration. Understand how charged molecules interact in agarose gel and learn to interpret the results through visible precipitation lines.
