Carrier particle Agglutination

Slides:



Advertisements
Similar presentations
Agglutination: Latex Agglutination
Advertisements

Part Three Basic Test Methods
Serology 1. Serology In vitro Antigen- Antibody reactions Antigen- Antibody reactions are classified according to the physical state of antigen into:
Agglutination reaction immunology Tutorial MLT402
Serological reactions in Microbiology Tatyana Ivakhnyuk The Department of Infectious Diseases and Epidemiology with Course of Microbiology, Virology and.
© 2004 Wadsworth – Thomson Learning Immunology Tutorial Introduction & Course outline By: Moh’d J. Al Khatatneh.
Immuoagglutination test Yasser M. alatawi Pharm.D College of Pharmacy Umm Al-Qura University.
detection of Rheumatoid factor by using LatexAgglutination
Principles of Immunology Antigen-Antibody Interactions 4/25/06
TREPONEMA,BORRELIA,LEPTOSPIR A Spirochetes. They are gram negative bacteria Long, thin, helical, and motile.
بسم الله الرحمن الرحيم.
Performance of Serological tests for Syphilis. Learning Objectives Know the rationale for performance of syphilis test in ICTC Know all the processes.
INTRODUCTION. Immune System The immune system is a system of biological structures and processes within an organism that protects against disease. processesorganismdisease.
Carrier particle Agglutination
Application of immunological tests
Agglutination tests HA & HI.
Antigen-antibody reaction
Done by: Bilal M. Marwa, Abdullah Al-Harby. From the slides of: Dr. Jad AlRab.
Introduction to Serology
LABORATORY DIAGNOSIS OF SYPHILIS
- Primary serological tests: (Marker techniques) e.g. Enzyme linked immuonosorbent assay (ELISA) Immuno flurescent antibody technique (IFAT) Radio immuno.
2. Basic Immunologic Procedures Part 5 Agglutination
Unit #6C – Clinical Laboratory Testing – Basic Serology/Immunology Cecile Sanders, M.Ed., MT(ASCP), CLS (NCA)
Complement Fixation Test
Overview of the Immune System Immune System Innate (Nonspecific) Cellular Components Humoral Components Acquired (Adaptive Specific) Cell-Mediated Humoral.
C - reactive protein. C - reactive protein ( CRP ) ◌ C-reactive protein was originally discovered as a substance in the serum of patients with acute inflammation.
R HEUMATOID F ACTOR ( RF ). Rheumatoid arthritis is a chronic inflammatory disorder that affects all joints either in hands or feet and other areas of.
Agglutination Definition – the clumping together of antigen bearing cells, microorganisms or particles in the presence of specific antibodies. Particles.
RHEUMATOID ARTHRITIS (RA)
Agglutination Reaction
Precipitation tests. Flocculation tests Countercurrent Immuno-electrophoreis Immunodiffusion Assay (ID) 1- VDRL. 2-RPR 3-One step H.pylori Test device.
© 2004 Wadsworth – Thomson Learning Chapter 19 Diagnostic Immunology.
Agglutination Part 2.
Indirect Haemagglutination
( Antistreptolysin O Titer )
C-Reactive Protein (CRP). CRP CRP is a serologic test which is used for diagnosis of Inflammations and Infections. CRP was so named because it was first.
The Antiglobulin Test ( Direct & Indirect )
Agglutination 1.
C- reactive protein (CRP)
Rheumatoid Factor (RF)
CRP C- reactive protein.
BIOLOGICAL EVIDENCE BLOOD.
Y +  AGGLUTINATION By: F.Mirzaalian.
Complement fixation test
Serological test M.Sc. Sarah Ahmed.
Antibody-Antigen Reactions
( Antistreptolysin O Titer )
Test for Rheumatoid Factors
Properties of Treponema pallidum
Rheumatoid Factor (RF)
The Immune System.
Agglutination test It is one of important laboratory method to detect antigen antibody reaction. It provides flexible and useful method for semi quantitating.
Enzyme-Linked Immunosorbent Assay [ELISA]
CRP C- reactive protein.
CLINICAL PATHOLOGY: C-Reactive Protein
ERYTHROCYTE SEDIMENTATION RATE (ESR)
Blood.
Coombs test practical(3)
AUTOIMMUNE DISEASES.
Methods for Ag-Ab detection
Part 3.
Unit #5C – Clinical Laboratory Testing – Basic Serology/Immunology
Unit #5C – Clinical Laboratory Testing – Basic Serology/Immunology
Blood group and cross matching
Antigen-Antibody reactions
( Antistreptolysin O Titer )
Chapter 12 Blood.
Chapter 17 Topics Phenotypic Genotypic Immunological Method.
( Antistreptolysin O Titer )
Presentation transcript:

Carrier particle Agglutination Latex agglutination flocculation

Carrier particle agglutination test Agglutination test: Qualitative/ Quantitative(Ab titer) agglutination test. It is used to determine Ag or Ab presence and amount(titer) Carrier particles: Latex, RBC’s, charcoal, protein A of Staph. Aureus. It is an indirect test. It is used to detect Ag or Ab in the patient sample. Latex particles are inert polystyrene substance(0.79-0.81mm).Ag or Ab attach spontaneously to it.

Latex Agglutination Test Rheumatoid arthritis (RA) Is traditionally considered a chronic, inflammatory autoimmune disorder that causes the immune system to attack the joints. It is a disabling and painful inflammatory condition, which can lead to substantial loss of mobility due to pain and joint destruction. RA is a systemic disease, often affecting extra- articular tissues throughout the body including the skin, blood vessels, heart, lungs, and muscles. About 60% of RA patients are unable to work 10 years after the onset of their disease.

The Rheumatoid arthritis immune response appears to be directed against multiple antigenic determinants on the gamma globulin molecule.( Anti-IgG) In the test kit, latex particles are coated with human gamma globulin IgG molecule. In the screening test, however, positive results may also be obtained due to others disorders such as: Systemic Lupus Erythematosis (SLE) Polyarteritis nodosa Dermatomyositis or Scleroderma. Some diseases such as Cirrhosis, Hepatitis,, Syphilis, Sub acute bacterial endocarditis and Lymphomas may also give rise to agglutination reactions. Titers in these conditions are usually less than 1:20

Method: Sample collection: Collect 5 – 10 ml venous blood into sterile tube without anticoagulant Allow to clot at room temp. Formation of clot must be complete Separate serum and store at 2-8 °C It is preferable to test samples within 2-3 days

Method: Screening test Bring all reagents to room temp. Prepare 1:20 dilution of test serum by adding 50 μl of serum to 1 ml of glycine buffer Place 50 μl of diluted serum on to the test slide Add 1 drop of well shaken latex reagent Mix the two drops together with a clean stirrer and spread out to the edge of the test area Rock the slide gently and observe for macro agglutination Read at 2 minutes under a direct light source A definite clumping is reported as reactive (R). No clumping is reported as non-reactive (N).

CRP Latex Kit C-reactive protein (CRP) is a non-specific test. It is used by a doctor to detect inflammation if there is a high suspicion of tissue injury or infection somewhere in the body, but the test cannot tell where the inflammation is or what condition is causing it. CRP is not diagnostic of any condition, but it can be used together with signs and symptoms and other tests to evaluate an individual for an acute or chronic inflammatory condition

CRP Latex Kit

CRP Latex Agglutination test The same principle of Latex kits . CRP is a protein present normally in human serum. Anti-CRP is coated on latex to detect CRP in patient sample to detect CRP in patient sample. CRP is used mainly as a marker of inflammation It may also be ordered on a regular basis to monitor conditions such as rheumatoid arthritis and Lupus and is often repeated at intervals to determine whether treatment is effective

CRP Test Elisa Kit Latex kit Latex Kit

Flocculation Test Rapid Plasma Reagin (RPR) Syphilis is a sexually transmitted disease caused by the spirochete Treponema pallidum. The RPR (Rapid Plasma Reagin) Card test is a presumptive serologic screening test for syphilis. The serum of a person with syphilis contains a non-specific anti-lipid antibody (termed Reagin), which is not found in normal serum.

Syphilis infection starts the breakdown of the patient's own tissue cells. Fatty substances which are released, combine with protein from Treponema pallidum to form an antigen which stimulates the body to produce antibodies against both the body's tissue lipids (non-specific or non-treponemal) as well as the T. pallidum protein (specific or treponemal). The RPR Card test detects the nonspecific antilipid antibody and is referred to as a non- treponemal test for syphilis.

Principle of the Method The RPR test is a non-treponemal slide agglutination test for the qualitative and semi-quantitative detection of plasma reagins in human serum. Carbon particles coated with a lipid complex are agglutinated when mixed with samples containing reagins.

The known RPR antigen consists of Cardiolipin, lecithin, and cholesterol bound to charcoal particles Charcoal makes the reaction visible. If the patient has syphilis, the antilipid antibodies (Reagin) in his/her serum will cross-react with the known RPR lipid antigens giving a visible clumping of the charcoal particles.

Procedure: Place 50 µl of sample and one drop of each +ve and –ve controls into separate circles on the test slide. Swirl the RPR-carbon reagent gently before using. Place the micropipette in a vertical position and perpendicular to the slide, and add one drop (20 µl) of this reagent next to the samples to be tested. Mix the drops with a stirrer, spreading them over the entire surface of the circle. Use different stirrers for each samples. Place the slide on a mechanical rotor (shaker) at 80-100 r.p.m. for 8 minutes. Read the test results immediately.

Read the results as follows: A definite clumping of the charcoal particles is reported as reactive (R) No clumping is reported as non-reactive (N).

Co-agglutination Protein A which is a cell wall component of Staphylococcus aureus is able to bind to Fc portion of most IgG Ab leaving Fab portion free to interact with Ag present in specimens. This test is used to detect Ag in specimens or isolated bacteria from cultures. Example: streptococcal serotyping, Bacterial Ag detection kit for meningitis.

Coated Ab on Staph protein A Co-agglutination It is an agglutination where Ab is coated on protein A of Staph. aureus . + Co-agglutination Y Y Y Y Y Y Y Y Y Y TEST Ag Coated Ab on Staph protein A