1. Online Counseling Resource YCMOU ELearning Drive…
Dec 18, 2007
Online Counseling Resource YCMOU ELearning Drive…
School of Architecture, Science and Technology
Yashwantrao Chavan Maharashtra Open University, Nashik – , India

2. OC-SBT052-CP4-03 Introduction
Dec 18, 2007
OC-SBT052-CP4-03 Introduction
Programmes and Courses
SEP–SBT052–Unit-04
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3. © 2008, YCMOU. All Rights Reserved.
Dec 18, 2007
Dec 18, 2007
Credits
Academic Inputs by
Arun Punaji More.
M.Sc. (Microbiology)‏
Experience: 11 Years
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4. How to Use This Resource
Dec 18, 2007
How to Use This Resource
Counselor at each study center should use this presentation to deliver lecture of minutes during Face-To-Face counseling.
Discussion about students difficulties or tutorial with assignments should follow the lecture for about minutes.
Handouts (with 6 slides on each A4 size page) of this presentation should be provided to each student.
Each student should discuss on the discussion forum all the terms which could not be understood. This will improve his writing skills and enhance knowledge level about topics, which shall be immensely useful for end exam.
Appear several times, for all the Self-Tests, available for this course.
Student can use handouts for last minutes preparation just before end exam.
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5. © 2008, YCMOU. All Rights Reserved.
Dec 18, 2007
Learning Objectives
After studying this module, you should be able to:
Describe the principle of complement fixation test
Describe the applications of complement fixation test
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6. © 2008, YCMOU. All Rights Reserved.
Dec 18, 2007
Introduction
Many soluble antigens of pathogenic microorganisms and viruses can not be detected by routinely used agglutination or precipitation test.
This happens because sometimes the antibodies formed in infections can not precipitate or agglutinate the antigens against which they are produced.
Therefore the diagnosis of such infection is not possible by agglutination or precipitation test; more sensitive serological test is required to diagnose such infections
By Complement fixation test one can detect 0.04mg of antibodies and 0.1 antigen; this module will show how it is possible.
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7. Complement Fixation Test-1
Dec 18, 2007
Complement Fixation Test-1
Complement fixation test utilize the capability of complement factor to fix to antigen-antibody complexes.
When antigen-antibody react forming the Ag-Ab complex, complement get activated and bind to these immune complexes resulting in the lysis of cells in the vicinity of immune complexes.
Complement fixation test is very sensitive and detect 0.04mg of antibody and 0.1mg of antigens of pathogenic agents.
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8. Complement Fixation Test-2
Dec 18, 2007
Complement Fixation Test-2
The complement fixation test is carried out in two stages:
In the first stage the known antigens solution and serum lacking complement are allowed to incubate for some time.
If the serum contain the corresponding antibodies against the known antigen they will react with each other.
This reaction mixture then is mixed with the freshly harvested complements from guinea pig serum and incubated for sometime.
During second stage, this incubated reaction mixture is mixed with indicator system consisting of sheep's RBS coated with Ab against them.
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9. Complement Fixation Test-3
Dec 18, 2007
Complement Fixation Test-3
If the desired antibodies are present in the serum then there will be formation of Ag-Ab complexes which will consume or fix the complements
During the second stage when sheep's RBS coated with Ab are added they will not be lysed as there is no free complement to bring about their lysis indicating the positive complement fixation test.
However if there are no antibodies against the desired antigen in the reaction mixture, complement will remain free to react with sheep' RBC and their Ab complexes bringing about the lysis of sheep's RBC indicating the negative complement fixation test.
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10. Complement Fixation Test-4
Dec 18, 2007
Complement Fixation Test-4
Wassermann test:-
The Wassermann test is based upon the principle of complement fixation test.
Wassermann test is utilized for the diagnosis of syphilis.
The Wassermann test consist of two stages; in the first stage the inactivated of patient's serum is incubated with Wassermann antigen and two units of guinea pig complement for one hour at 37°C.
In the second stage this reaction mixture is incubated with sheep's RBC coated with Ab against them.
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11. Complement Fixation Test-5
Dec 18, 2007
Complement Fixation Test-5
Wassermann test:-
If the antibodies against Treponema pallidum is present in the serum there will be Ag-Ab reaction which will fixed or consume the complement factors.
Therefore during the second stage when the sheep's RBC coated with antibodies are added, there is no lysis of these RBC indicating the positive Wassermann test.
In this test positive test control of Ag-Ab and complement are run in parallel.
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12. Complement Fixation Test-6
Dec 18, 2007
Complement Fixation Test-6
Ab titre estimation by complement fixation test:-
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13. Complement Fixation Test-7
Dec 18, 2007
Complement Fixation Test-7
Applications of CFT:-
To detect the antibodies against viruses in patients' serum to diagnose those viral infections.
To detect the antibodies against pathogenic fungi.
To detect the antibodies against Treponema pallidum in syphillus.
To detect the antibodies against the mycoplasma.
CTF is used to detect Ab against non hemagglutinating viruses.
CFT is used to detect IgG and IgM against number of infectious agents.
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14. Viral Hemagglutionation Inhibition Test-1
Dec 18, 2007
Viral Hemagglutionation Inhibition Test-1
Mechanism:-
Some viruses such as influenza, measles, mumps, mononucleosis can bind to R.B.C. And bring about their agglutination.
This agglutination of R.B.C. Is called as hemagglutination.
In this test, test serum, standard Ag of a virus and washed R.B.C. are mixed in a test tube, if Ab against the virus is present in the serum it will neutralize the virus and virus will not bind to R.B.C. and there will be no hemagglutination indicating positive test for the diagnosis of the infection caused by that virus.
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15. Applications of Hemagglutination Inhibition Test
Dec 18, 2007
Applications of Hemagglutination Inhibition Test
To detect the antibodies against measles, mumps, influenza and mononucleosis viruses.
To determine the titre of antibodies against these viruses.
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16. © 2008, YCMOU. All Rights Reserved.
Dec 18, 2007
What We Learn…..
Complement fixation test
Principle of complement fixation test
Serological applications of complement fixation test.
Viral hemagglutination inhibition test and its applications.
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17. Critical Thinking Question
Dec 18, 2007
Critical Thinking Question
Why CFT is not carried out in the infections caused by influenza, measles and mumps viruses routinely?
Why the test serum is incubated at 56°C for 30 minutes before conducting CFT?
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18. Hints For Critical Thinking Question
Dec 18, 2007
Hints For Critical Thinking Question
Measles, mumps and influenza are hemagglutinating viruses.
Comparison of CFT and viral hemagglutination inhibition test in terms of cost, complicity.
Serum contain inactive complements proteins.
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19. © 2008, YCMOU. All Rights Reserved.
Dec 18, 2007
Study Tips
Book
Title: Immunology
Author: Richard A. Goldsby, Thomas Kindt, Barbara A. Osborne.
Publication: W.H. Freeman and Company, New York.
Title: Textbook of Microbiology
Author: R. Ananthnarayan and C.K.J. paniker
Publication: Orient Longman private limited, Chennai.
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20. End of the Presentation
Dec 18, 2007
End of the Presentation
Thank You !
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