visible expression of the aggregation or clumping of antigens and antibodies

antigens or antibodies are absorbed onto the surface of particles known as carriers

latex particles

collodial charcoal

rbc

baterial cells

antibody molecules are bound to the latex beads

results:normal values reveal no agglutination

q.c: postive and negative values; dont use after expiration

clinical significance: c-reactive protein

rubella antibody

IgG and IgM R.A.

neutralization of toxins protect against circulating bacteria and viruses

complement causes hemolysis through a complex process known as the complement cascade

hemolysis is the lysis of rbcs

concentration or strength of an antibody

expressed as the highest dilution of the serum that procedures aggultination

temperature (IgM 4-27C and IgG 30-37C)

incubation time

pH

antigen/antibody ratio

double immunodiffusion test method

precipitation method

identity: precipitin band forms a single smooth arc.

non-identity: precipitin lines cross each other

partial identity: precipitin lines merge in a spur formation

combines diffusion with electrophoresis to speed up or sharpen results

electrophoresis separates molecules according to differences in their electric charge when they are placed in an electric field.

an electrical current is added to help antigens and antibodies move toward each other more quickly than simple diffusion

antibodies: positively charged migrate to negative electrode

antigens: negatively charged migrate to positive electrode

q.c.: positive and negative controls should be used; follow manufacters instructions

Clinical app: anitinuclear ribonucleoprotein; radioimmunoprecipitation

principle: antigens are electrophoresed in an antibody containing gel ; precipitate band forms a rocket

q.c.: control sera should be used; follow manufacture instructions

principle:a two stage procedure using agarose gel protein electrophoresis followed by immunoprecipitation

clinical applications: used primarily for identification of monoclonal immunoglobulins

relies on the principle of comeptitive binding. the analyte being detected competes with a radiolabed analyte for a limited number of binding sites on a high affinity antibody.

q.c.: control sera; manufacturer's instructions

clinical app: hepatits A IgM antibody; hepatitis A antigen

same method as RIA uses but uses two monoclonial antibodies of high affinity

advantages: faster rate; increased sensitivity

the luminescence in which the light emission is caused by the products of a specific chemical reaction

advantages: excellent sensitivity and dynamic range; doesn't require sample radiation

sample antigen binds to an antibody fixed onto a solid phase. a second antibody labeled with chemiluminescent label binds to the antigen-antibody complex on the solid phase

clincial app: hepatitis A IgM anitbody; hepatitis A antigen

naturally occuring molecules that catalyze certain biochemical reactions

react with suitable substrates to produce breakdown products that may be chromogenic, flurogenic, or luminescent

alkaline phosphatase

horseradish peroxidase

beta-galactosidase

principle:a plastic bead or plastic plate is coated with antigen. the antigen reacts with antibody in the patient serum.; most commonly used enzymes are peroxidase and alkaline phosphatase

q.c.: control sera; manufacturer's instructions

used for both antigen and antibody identification

principle: antibody conjugated with a fluorescent tag is added directly to unknown antigen that is fixed to a microscope slide; incubate and wash; antigens are typically visualized as bright apple green or orange-yellow objects against a dark background

q.c.: manufactorer's instructions

clincal app: cmv, hepatitis b, epstein-barr virus, rsv

patient's serum is diluted and placed on the slide to cover the antigen source.

q.c.:manufacturer's instructions

clincal app: sereological method most widely used to detect diverse antibodies

screening test ANA's

antigen substrate from tissue containing nuclei is fixed to a slide

most common tissues used are mouse liver or kidney, or cell-cultured fibroblasts grown on slides

presence of ana antibodies:

can indicate various sstemic autoimmune connective tissue disorders (hallmark of SLE)

often detected in liver diseases associated with autoimmunity such as activate hepatitis and primarily biliary cirrhois

can be seen in small percentages in the elderly w/o disease an dmall percentages in healthy nonelderly individuals

a classic method for demonstrating the presence of a complement fixing antibody in serum

varicella-zoster virus

herpes simplex infection

dna

rna

mostly widely used amplifications technique

produce millions of copies of a specific DNA sequence in approximately two hrs

pcr: human papiloma virus; coronary artery disease

rt-pcr: human immunodeficiency virus

principle: a combination of fluid dynamics, optic, laser science, high speed computers and fluid flurochrome- conjugated monoclonal antibodies

rapidly classify groups of cells within heterogeneous mixtures

cells are stained with flurochrome for specific reactivity

identification of cells

cell sorting before further analysis

principle: designed to perform microparticle enzyme immunoassay and ion capture; allows for both high and low molecular weight analytes to be measured

clinical application: endocrine function and fertility; cancer, hepatitis, and transplant

negative- no aggregates

mixed field- few isolated aggregates

weak (+/-) tiny aggregates barely visible turbid reddish supernatant

1+: a few small aggregates just visibly macroscopically

2+: medium sized aggregates clear supernatant

3+: several large aggregates clear supernatant

4+: all erythrocytes are combined into one solid aggregate clear supernatant

principle: latex agglutination test in which latex particles are coated with streptolysin O antigen

visible agglutinations seen

clincal app: elevated titer + for group A strep; strep skin infections produce low ASO titers; low titers can exist in healthy persons due to frequency of strep infections

principle: test for antibody detection based on interaction of soluble antigen with antibody; particles are macroscopically or microscopally visible

q.c.: manufacturer's instructions

clincal app: syphilis VDRL/ RPR

principal: detects antibodies to rbc antigens

q.c.: manufacturer's instructions

clincal app: ABO blood typing, Rh antigens, Hepatitis B/C, HIV I/II