Immuno Lab

measures of antibody binding strength, and are important characteristics of the immune response

The strength of a single antibody-antigen interaction. ex: antibodies with high affinity bind more strongly and stably.

the total strength of all interactions between an antibody and an antigen.

avidity is always greater than affinity.

# binding sites, structural arrangements

2 binding sites

10 binding sites

2 binding sites

IgG

IgG

the test's ability to correctly identify those with antibodies (true positives)

the test's ability to identify those without the antibodies (true negative)

the test correctly identified 25 out of 50 patient with the disease, and all 50 healthy people.

this means that the test correctly identified all 50 patients with the disease, but only 25 out of the 50 healthy people.

this means that 90% of people with the disease tested positive, and 80% of people without the disease tested negative.

both sensitivity and specificity

Confirming a disease if a test result is positive, but it's still possible for people to get a negative test result when they actually have the disease.

Sharpes
Biohazard
Trace Chemo
RCRA Hazard
Pharmaceutical
Radioactive

Primary and secondary syphillius --diagnosed by presence of T. pallidum in exudates of skin lesions

dark condensor - light need to do ASAP

flueorescent labeled Ab to conjugate to T. pallidum

no active lesions, may be secondary or teritary syphillis - need to test for Abs.

determine presence of Ab (reagin) that forms against cardiolipin, lipid material released in damaged cells.

specific types of precipitation that occurs over a narrow range of antigen concentrations.

modified VDRL test involving agglutination; The cardiolipin -- containing antigen suspension is bound to charcoal particles. Check to titer.

Detects antibodies to T. pallidium or specific antigens; positive nontreponemal (screening) must be confirmed.

uses slide with green gluorescent labeled antibody - for positive result (shows Ab present)

colored gelatin particles to coated with trep antigens

occurs when antigens are found naturally on a particle

indicates presence of patient antibody

employs particles that are COATED with antigens not normally found on their surfaces

indicates presence of patient antibody

can detect antibodies to viruses

antibdoy rather than antigen is attached to carrier particle. Particles coated with reagent antibody.

agglutination indicates presence of patient antibody.

haptens attached to carrier particles. Rxn based on inhibition of agglutination due to competition with a soluble antibody.

POSITIVE test - indicating presence of patient ANTIGEN

reaction uses same principle as agglutination inhibition except RBCs are indicator particles

POSITIVE test - indicating presence of patient ANTIBODY

written plan to provide necessary protection, free of charge.
Components to plan:
1. Engineering Controls - sharps disposal etc.
2. Work Practice Controls - standard precautions
3. Personal Protective Equipment
4. Medical - hep. B vaccine free of charge
5. Documentation - annual training

collection of microorganisms on the human body

no apparent benefit or harm to either

both humans and bacteria benefit from each other

bacteria result in harm to the host

excess of antibody, suboptimal ppt. too many Abs for lattice formation

optimal ratio of antibody to antigen, results in optimal ppt.

excess of antigen, suboptimal ppt.

PPT involves SOLUBLE antigens, while
agglutination involves PARTICULATE antigens

formation of visible solid ppt when soluble antigens bind to their specific antibodies

clumping together of particulate antigens (like cells) due to binding with specific antibodies, essentially creating clumps instead of ppt.

UREASE

Produces protein CagA and a vacuolating cytotoxin, or VacA

Enzyme that provides a buffer zone around the bacteria that protects it from the effects of stomach acid.

Detect urease from stomach biopsy
CLO detects urease activity in gastric mucosal biopsies - gell turns HOT pink due to increase in pH
Urea Breath Test

Endotoxin (lipid A) is found in the LPS layer of cell wall of gram
Exotoxin are protein moleucles released from the living bacteria

lipid A

a toxin found in the LPS (lipopolysaccharide) layer of cell wall of gram - bacteria.
When cell dies - it releases the Lipid A in host
Causes host response to release cytokines, TNF, and platelet activating factor.

specific and targeted activity.
Protein molecules released from living bacteria (mostly gram + bacteria) and considered most potent molecules known to harm living moleulces.
Neurotoxins, cytotoxins, enterotoxins
Exotoxins bind to specific receptors on host cells that activate specific activity of a toxin.

1. sharps
2. biohazards
3. pathological

piercing skin biohazard waste

anything contaminated with blood or biohazard material

tissues organs, fluids, and body parts

proportion of people who DO NOT have the disease and who have a negative test

True positives / (true positives + false negatives)

True negatives / (true negatives + false positives)

uses the calculated standard deviation (or known SD)

California dept of health, Laboratory Field Service (STATE LEVEL)

May remain positive for years, which decreases the specifity of these tests confirming active or eradicated infection.

One of the most common pathogenic bacteria and causes a variety of diseases

the major virulence factor of GAS has a net negative charge at the amino-terminal end that helps inhibit phagocytosis
-limits deposition of C3 on the bacterial surface, diminishing complement.

Capture assay

1. Sandwich immunoassay
2. Ab (rather than ag) is bound to the solid phase during incubation
3. Enzyme labeled Ab added and attaches to available patient Ab
4. Sandwich is formed
5. Enzyme activity directly proportional to amount of Ab.

Nontreponemal tests first (RPR) or VDRL first (screening test)
if positive nontrep test then Treponemal tests

Ab against T. pallidium or antigens

1. Fluorescent treponemal antibody absorption (FTA-ABS)
2. Agglutination Tests (TP-PA)

HIV screening tests are known to have false positives. Specimens with some bands, but do not meet criteria. REPEAT WB AGAIN; IF STILL INDETERMINATE THEN REPEAT WB AGAIN IN ABOUT 4-6 WEEKS USING A NEW SPECIMEN

confirmatory test for a positive HIV Ab screen test (ELISA)

Utilizes nitrocellulose or nylon strips contrining individaul HIV proteins that separate gel electrophoeses and blotted onto test.

antibodies move across strip.

the strength of a single bond or interaction between two molecules at a specific binding site.

the total strenght of all non-covalent interactions between 2 species that interact multivalently . Avidity takes into account multiple binding sites or interactions.

antibodies with fewer binding sites have high affinity and low avidity.

agglutination reactions involving RBCs

patient RBCs are mixed with antisera of IgM type can be used to determine the presence of absence of the A and B antigens.

rapid covid, and pregnancy test

1. Patient sample is added to a cassette containing antibody labeled with colodial gold.
2. Sample combines with antibody and is moved along by capillary flow.
3. Monoclonal antibody to the analyte captures the patient antigen attached to gold-labeled antibody.
4. Control line has antibody tha captures collodial gold-labeled antibody

IgM