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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-CP4-01 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: Define affinity, avidity and specificity of antibody Describe various methods of agglutination reaction Describe applications of agglutination reaction in diagnosis of diseases.

6 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.6 Introduction  When immune system mount an immunological response to an invading pathogen or its toxic product, effector immune cells are produced to kill invading pathogen and antibodies are produced to neutralize the pathogen and its toxic product.  Since the antibodies have got specificity with its corresponding antigen, antigen antibody reaction in vitro is used to diagnose an infection.  In this module the students shall learn the underlining principle of diagnosis involving Ag-Ab reaction in vitro.

7 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.7 Nature of Antibody-Antigen Reaction-1  The antibody react with its corresponding antigen at optimum pH and electrolyte concentration in reversible manner.  Antibody react with its corresponding antigen which has stimulated its production very specifically.  The antigen antibody combining involve following week forces: Hydrogen bonds Electrostatic bonds Hydrophobic bonds Van der Waal forces

8 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.8 Nature of Antibody-Antigen Reaction-2  Each antigen-antibody reaction is unique in its following properties Affinity of Ag-Ab reaction Avidity of Ag-Ab reaction Specificity of Ag-Ab reaction

9 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.9 Nature of Antibody-Antigen Reaction-3  Affinity of Ag-Ab reaction:- Strength between single antigen determinant and a single antibody combining site is expressed in term of affinity of Ag-Ab reaction. Affinity of Ag-Ab is also product of attracting and repulsive forces between Antigen and antibody.

10 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.10 Nature of Antibody-Antigen Reaction-4  Avidity of Ag-Ab reaction:- The total overall strength between antigen and antibody combining is termed as avidity of antigen antibody reaction. The avidity is dependent on the valency of antigen as well as valency of antibody.

11 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.11 Nature of Antibody-Antigen Reaction-5  Affinity and avidity of Ag-Ab reaction:- High affinity means interaction or association of antigen and antibody remain stable thus can be easily detected in vitro. High avidity means interaction or association of antigen and antibody remain stable thus can be easily detected in vitro.

12 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.12 Nature of Antibody-Antigen Reaction-6  Particulate antigens when react with antibodies at equivalent concentration, particulate antigens get coagulated forming visible clumps of particulate antigens; this clumping of particulate antigens is called as agglutination of antigens.  When pathogenic organisms enter into the body antibodies are produced against number of surface antigens of the pathogens.  When the serum containing the antibodies against such surface antigens is mixed with the culture suspension of the pathogen, the antigens get cross linked by the antibodies resulting in the agglutination of the pathogens.

13 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.13 Agglutination-1 The clumping formation of the particulate antigens is explained with lattice of antigen formation when the antigen molecules get cross linked at the equivalent concentration of antigens and antibodies.

14 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.14 Agglutination-Test-1  The agglutination is carried out in number of ways and accordingly there are different types of agglutination tests Direct agglutination test Indirect agglutination (or passive agglutination test) ‏ Haemagglutination Haemagglutination inhibition test Coombs' test

15 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.15 Agglutination-Test-2  Direct agglutination test:- The direct agglutination test is carried out to detect particulate antigens such as red blood cells, bacteria and fungi by serum antibodies. In this test fixed amount of known particulate antigen solution is mixed with different solution of serum antibodies having serially diluted twofold times to determine the relative titre of the antibodies in the serum.

16 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.16 Agglutination-Test-3  Passive agglutination test:- Very microscopic antigens such as originated from viruses though crossed linked they do not form visible agglutination; Therefore such antigens are bound to some larger inert particles and are subjected to agglutination test. Here the micro latex particles are coated with such antigens of infectious agents and are mixed with anti-sera which bring about visible agglutination of these latex particles making the test visible.

17 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.17 Agglutination-Test-4  Haemagglutination test:- The antibodies against the surface antigens of red blood cells result into cross linking of the cells resulting in the agglutination which is called as haemagglutination. Haemagglutination is used to determine blood types or blood groups.

18 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.18 Agglutination-Test-5  Haemagglutination inhibition test:- Like antibodies against the surface antigens of red blood cells cause agglutination of these cells, certain viruses are also having capability of bringing about the agglutination of red blood cells. To diagnose the infections caused by such viruses haemagglutination inhibition test is carried out. The viruses which cause haemagglutination and are diagnosed by haemagglutination inhibition test are influenza, measles, mononucleosis, mumps viruses, etc.

19 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.19 Agglutination-Test-6  Haemagglutination inhibition test:- In this test anti-sera is mixed with fresh red blood cells and then with suspected virion, if antibodies against these suspected viruses are present the antibodies against them will neutralize them and therefore they can not bring the haemolysis of red blood cells thus inhibition of haemagglutination is positive indication of particular viral infection.

20 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.20 Agglutination-Test-7  Coomb's test:- This is modified agglutination test. Some antibodies can not agglutinate the particulate antigen though they react with the antigens. Such antibodies are called as non agglutinating antibodies. Therefore the antibodies against such non agglutinating antibodies are produced in rabbit; these are called as anti- immunoglobulins. These anti-immunoglobulins when added into the antigen and antibody mixture, bring about the agglutination of the particulate antigens.

21 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.21 Agglutination-Test-8  Coomb's test:- The Coomb's test is used to detect the antibodies against the Rh antigen on red blood cells in mother's blood. Coomb's test is routinely used to detect Rh incompatibility. In the indirect Coomb's test Rh antibodies are detected in Rh negative mother's blood. Whereas the direct Coomb's test detect the Rh positive red blood cells of Rh positive fetus.

22 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.22 Agglutination-Test-9  Direct Coomb's test:- + ↔ Patient’s RBCs Coombs Reagent (Antiglobulin)‏

23 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.23 Agglutination-Test-10  Indirect Coomb's test:- Patient’s Serum Target RBCs + ↔ Step 1 + ↔ Coombs Reagent (Antiglobulin)‏ Step 2

24 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.24 What We Learn…..  Various methods of agglutination reactions.  Mechanisms of agglutination reactions.  Applications of agglutination reactions.  Coomb's test and its applications.  Haemagglutination inhibition test.

25 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.25 Critical Thinking Question  Why agglutination reactions are more widely used over the other serological reactions in bacterial infections?

26 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.26 Hints For Critical Thinking Question  Particulate antigens  Specificity  Sensitivity  Reliability  Time of test

27 School of Science and Technology, Online Counseling Resource… © 2008, YCMOU. All Rights Reserved.27 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. Anathanarayan & C.K.J. Paniker Publication: Publication: Orient Longman publication, Chennai.

28 © 2008, YCMOU. All Rights Reserved.28 End of the Presentation Thank You !


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