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Online Counseling Resource YCMOU ELearning Drive… School of Architecture, Science and Technology Yashwantrao Chavan Maharashtra Open University, Nashik.

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Presentation on theme: "Online Counseling Resource YCMOU ELearning Drive… School of Architecture, Science and Technology Yashwantrao Chavan Maharashtra Open University, Nashik."— Presentation transcript:

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

2 © 2008, YCMOU. All Rights Reserved.2 OC-SBT052-CP4-03 Introduction Programmes and Courses SEP–SBT052–Unit-04

3 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.3 Credits Academic Inputs by Arun Punaji More. M.Sc. (Microbiology) Experience: 11 Years

4 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.4 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.

5 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.5 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

6 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.6 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.

7 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.7 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.

8 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.8 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.

9 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.9 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.

10 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.10 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.

11 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.11 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.

12 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.12 Complement Fixation Test-6 Ab titre estimation by complement fixation test:-

13 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.13 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.

14 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.14 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.

15 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.15 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.

16 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.16 What We Learn….. Complement fixation test Principle of complement fixation test Serological applications of complement fixation test. Viral hemagglutination inhibition test and its applications.

17 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.17 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?

18 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.18 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.

19 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.19 Study Tips Book Title: Immunology Title: Immunology Author: Author: Richard A. Goldsby, Thomas Kindt, Barbara A. Osborne. Publication: Publication: W.H. Freeman and Company, New York. Book Title: Textbook of Microbiology Author: Author: R. Ananthnarayan and C.K.J. paniker Publication: Publication: Orient Longman private limited, Chennai.

20 © 2008, YCMOU. All Rights Reserved.20 End of the Presentation Thank You !


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