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94 Cards in this Set

  • Front
  • Back
The purpose of vaccines is to stimulate active immunity and to create an immune memory so that exposure to the active disease microorganism will stimulate an already primed immune system to fight the disease.
The purpose of vaccines is to stimulate _____ and to create an _____ so that exposure to the _____ will stimulate an already primed _____ to fight the disease.
The Center for Biologics Evaluation and Research, a department within the FDA, approve and monitor vaccinations.
The _____ , a department within the _____ , approve and monitor vaccinations.
Herd immunity theory is a form of immunity that occurs when the vaccination of a significant portion of a population, or herd, provides a measure of protection for individuals who have not developed immunity. The rate for herd immunity is usually 80-85%, but differs depending on the disease.
_____ is a form of immunity that occurs when the vaccination of a _____ of a population, or herd, provides a measure of protection for individuals who have not developed _____. The rate for herd immunity is usually _____ %, but differs depending on the _____.
In 1000 BC, Chinese 'immunized' themselves by inhaling dried powders made from the crusts of smallpox lesions.

In 1798 AD, Edward Jenner used cross reactivity of cow pox to prevent smallpox.

In 1880 AD, Louis Pasteur used attenuated vaccinations, which use live viruses with decreased virulence. He injected chickens with old cultures which caused the development of antibodies but not the disease.
In _____, Chinese 'immunized' themselves by inhaling _____ made from the crusts of _____.

In _____, _____ used cross reactivity of _____ to prevent smallpox.

In 1880 AD, Louis Pasteur used attenuated vaccinations, which use live viruses with decreased virulence. He injected chickens with old cultures which caused the development of antibodies but not the disease.
Types of Vaccinations:
Cross reactive (rarely used today)
Attenuated (MMR)
Inactive (flu shot)
Synthetic
Types of Vaccinations:
_____
_____
_____
_____
Types of Synthetic Vaccinations:
subunit - components of pathogens are injected with adjuvants
conjugated - protein of pathogen is conjugated with bacterial polysaccharide to create greater immune response
naked DNA - produces immunity to genes of pathogens
Types of Synthetic Vaccinations:
_____ - _____ of pathogens are injected with _____
_____ - protein of pathogen is _____ with bacterial polysaccharide to create greater _____
_____ - produces immunity to _____ of pathogens
FDA Requirements for a vaccine:
Produce protective immunity with only minimal side effects.
Be immunogenic enough to produce a strong and measurable immune response.
Be stable during its shelf-life, with potency remaining at a proper level.
FDA Requirements for a vaccine:
Produce _____ with only minimal _____
Be _____ enough to produce a _____ and _____ immune response.
Be _____ during its _____, with _____ remaining at a proper level.
The innate immune response to a vaccine occurs within minutes or hours of the vaccination. Dendritic cells can sense components of bacterial, viruses, parasites, and fungi through pathogen-recognition receptors (Toll-like receptors). This trigger naive antigen-specific T-cells, which launches the adaptive immune response.

Vaccine-induced immunity can wane. A second dose of vaccine could improve protection.
The _____ immune response to a vaccine occurs within _____ or _____ of the vaccination. _____ can sense components of bacterial, viruses, parasites, and fungi through _____ receptors (_____). This trigger naive _____, which launches the _____ immune response.

_____ immunity can wane. A _____ of vaccine could improve protection.
Production of a monoclonal antibody
develop an antigen
inject animal with antigen, usually with adjuvant, several booster shots will be required to increase titer
harvest antibodies from animal
spleen cells are mixed with myeloma cells and an agent such as polyethylalne glycol to cause the cells to randomly fuse together
selective media is used to obtain only myeloma/spleen hybridoma cells
hybrid cells are grown into hybridomas (clones) which are then moved to its only well to grow
specific cells with antibody of choice can then be centrifuged and separated to be use at well

Takes about one year for the whole process.
Production of a monoclonal antibody
develop an _____
inject animal with _____, usually with _____, several _____ will be required to increase _____
_____ antibodies from animal
_____ are mixed with myeloma cells and an agent such as _____ to cause the cells to randomly _____
_____ is used to obtain only myeloma/spleen _____ cells
_____ cells are grown into _____ (_____) which are then moved to its own _____ to grow
specific cells with _____ of choice can then be _____ and _____ to be used at well

Takes about _____ for the whole process.
Monoclonal antibodies can be used in clinical diagnosis, treatment, and research.
_____ antibodies can be used in clinical diagnosis, treatment, and research.
CLSI stands for Clinical and Laboratory Standards Institute.
CLSI stands for _____.
Serology use red topped tubes.

Blood should be allowed to clot and the serum removed promptly afterward.

Store at 4C for up to 72 hours, if more than 72 hours store at -20C

Compliment can be deactivated by heating the serum at 56C for 30 minutes.
Serology use _____ topped tubes.

Blood should be allowed to _____ and the _____ removed promptly afterward.

Store at _____ for up to _____ hours, if more than _____ hours store at _____.

_____ can be deactivated by heating the serum at _____ for _____ minutes.
To separate the serum in blood:
Allow blood to sit at room temperature for 1 hour.
Loosen the clot from the sides of the tubes
Centrifuge for 10 min at moderate speed
Serum should be transferred to clean tube unless gel is present
Do not allow serum to get contaminated with erythrocytes when pulling it off the clot
Seal the serum tube.
To separate the serum in blood:
Allow blood to sit at _____ for _____ hour.
Loosen the _____ from the sides of the tubes
_____ for _____ min at moderate speed
Serum should be transferred to a _____ unless gel is present
Do not allow _____ to get contaminated with _____ when pulling it off the _____
Seal the _____ tube.
Serology specimens that can be tested include:
serum
urine
CSF
other body fluids
Serology specimens that can be tested include:
_____
_____
_____
_____
Dilutions are used to make a weaker solution from a stronger one.

Dilutions are expressed as a ratio.

The dilution factor is the reciprocal of the dilution made. It is used to correct for having used a diluted sample in a determination rather than the undiluted sample.

Serial dilutions is a dilution series in which all dilutions including or following the first one, are the same.
_____ are used to make a _____ solution from a _____ one.

_____ are expressed as a ratio.

The _____ is the _____ of the dilution made. It is used to correct for having used a _____ sample in a _____ rather than the _____ sample.

_____ is a dilution series in which all dilutions _____ or _____ the first one, are the _____.
Antibody titer is the reciprocal of the highest dilution of the patient's serum in which the antibody is still detectable.
Antibody _____ is the reciprocal of the _____ dilution of the patient's serum in which the _____ is still detectable.
Point of Care Testing is a laboratory assay performed near the patient. It can provide results very quickly but can be costly and difficult to maintain quality control and quality assurance.
_____ is a laboratory assay performed near the patient. It can provide results _____ but can be _____ and _____ to maintain quality control and _____.
CLIA '88 tests
Waived - simple tests anyone can do (POCT, home pregnancy)
Moderately complex - automated tests (MLTs)
Highly complex - (MLS, require judgement)
Provider-performed microscopy - (Doctor)
CLIA '88 tests
_____ - simple tests anyone can do (POCT, home pregnancy)
_____ - automated tests (MLTs)
_____ - (MLS, require judgement)
_____ - (Doctor)
Quality control standards use positive and negative controls. Samples sent to the lab and run and then sent back to check for lab accuracy. 5-10 samples are required to keep accreditation.
_____ standards use _____ and _____ controls. Samples sent to the _____ and run and then sent _____ to check for lab accuracy. _____ samples are required to keep _____.
Precipitation and agglutination reactions were the first immunoassays developed and rely on the fact that antibodies and most antigens have multiple binding sites which result in the formation of large complexes when combined at the appropriate concentrations.

These assays are not very sensitive because a positive reaction of antibody and antigen is actually seen unamplified by the human eye. Using optics to register the differences between uncomplexed and complexed antigen and antibody results in some improvement of the sensitivity.

Two optical techniques are turbidometry and nephelometry.
_____ and _____ reactions were the first immunoassays developed and rely on the fact that _____ and most _____ have _____ binding sites which result in the formation of _____ when combined at the appropriate _____.

These assays are _____ because a positive reaction of antibody and antigen is actually seen _____ by the human eye. Using _____ to register the differences between _____ and _____ antigen and antibody results in some _____ of the sensitivity.

Two optical techniques are _____ and _____ .
Precipitation is the cross-linking of a soluble antigen to create an insoluble precipitate that is visible.
_____ is the cross-linking of a _____ antigen to create an _____ precipitate that is _____.
Agglutination is the cross-linking of particulate antigens (bacteria, cells, or latex particles) to form larger complexes that are also visible.
_____ is the cross-linking of _____ antigens (bacteria, cells, or latex particles) to form _____ that are also _____.
All serologic reactions, including unlabeled or labeled, rely on affinity, avidity, and specificity of the antibody to bind to the antigen.

The higher the affinity, the more sensitive is the reaction because more of the antigen and antibody will be in complexed form.

IgM has a weaker affinity than IgG because IgG is usually produced after the somatic mutational events that improve affinity.

Steric hindrances can also affect the binding when the antigens bound are big enough to block binding by the other binding sites of the immunoglobulin.
All _____ reactions, including _____ or _____ , rely on _____, _____, and _____ of the antibody to bind to the antigen.

The higher the _____, the more _____ is the reaction because more of the antigen and antibody will be in _____ form.

_____ has a weaker affinity than _____ because _____ is usually produced after the _____ mutational events that improve _____ .

_____ can also affect the _____ when the antigens bound are big enough to _____ binding by the other _____ of the _____.
Avidity is the number of binding sites on the immunoglobulin times the affinity.
_____ is the number of _____ on the immunoglobulin times the _____.
Affinity is a measure of the strength of the binding of 1 Fab region with its corresponding epitope on the antigen.
_____ is a measure of the strength of the _____ of 1 Fab region with its corresponding _____ on the antigen.
Equivalence is the part of the precipitation curve when the number of paratopes is approximately equal to the number of epitopes.
_____ is the part of the precipitation curve when the number of _____ is approximately equal to the number of _____.
Prozone is the part of the precipitation curve when the number of antibody molecules are greater than the number of antigens and there is no need for bridging of two antigen molecules by an antibody molecule causing little or no precipitation.
_____ is the part of the precipitation curve when the number of _____ molecules are greater than the number of _____ and there is no need for _____ of two antigen molecules by an antibody molecule causing little or no _____.
Postzone is the part of the precipitation curve when the antigen molecules are greater than the number of antibody molecules, causing two antigen molecules to bind to one antibody and there is no bridging of two antibody molecules causing little or no precipitation.
_____ is the part of the precipitation curve when the _____ molecules are greater than the number of _____ molecules, causing two antigen molecules to bind to one antibody and there is no bridging of two antibody molecules causing little or no _____.
In the prozone and postzone situation, the patient would appear negative for antibody to the tested antigen.

In a prozone situation, the serum can be diluted and tested again to reach equivalence, while in the postzone situation, blood can be drawn again in 1 or 2 weeks after the patient has formed more antibodies to reach equivalence.
In the prozone and postzone situation, the patient would appear negative for antibody to the tested antigen.

In a _____ situation, the serum can be _____ and tested again to reach _____, while in the _____ situation, blood can be drawn again in _____ after the patient has formed more _____ to reach _____.
Immunodiffusion techniques measure precipitation reactions.

Precipitation is a very easy technique used to detect antigen-antibody interactions. Antigens and antibodies are placed in different wells in an agar plate and allowed to diffuse. The resulting pattern of precipitation is then analyzed.

The measuring limit of precipitation is approximately 20 ug/mL of antibody and antigen.
_____ techniques measure precipitation reactions.

_____ is a very easy technique used to detect _____ interactions. _____ and _____ are placed in different wells in an _____ and allowed to _____. The resulting _____ of precipitation is then analyzed.

The measuring _____ of precipitation is approximately _____ of antibody and antigen.
Precipitation is the least sensitive serological technique because it requires the visualization of two substances in solution as they come out of solution. It is not amplified by the antigen being on a particle, nor is it amplified by any methods to link this reaction to another more visible reaction.

Immunodiffusion is a clinically important assay for fungal antigens and for some research purposes because it can give information concerning the relatedness of antigens and the minimum number off antigen and antibody pairs, which is difficult to garner in any other way.
_____ is the least sensitive _____ technique because it requires the _____ of two substances in solution as they come out of _____. It is _____ by the antigen being on a particle, nor is it _____ by any methods to link this _____ to another _____ visible reaction.

_____ is a clinically important assay for _____ and for some research purposes because it can give _____ concerning the _____ of antigens and the minimum number of _____ and _____ pairs, which is difficult to garner in any other way.
Two examples of immunodiffusion techniques are:
double diffusion gel precipitation (Ouchterlony, immunodiffusion reactions)
single diffusion precipitation (radial immunodiffusion)
Two examples of immunodiffusion techniques are:
_____
_____
Ouchterlony is also called double diffusion gel precipitation or immunodiffusion (ID) reactions and is a qualitative procedure.

Both the antibody and antigen diffuse through agar or agarose. As they diffuse their concentrations diminish and when the antibody and antigen reach equivalence, they form a precipitin line.

The precipitin line is formed by the lattice structure (cross-linked) formed by the antigen and antibody complexes.
_____ is also called double diffusion gel precipitation or _____ (_____) reactions and is a _____ procedure.

Both the _____ and _____ diffuse through _____ or _____. As they _____ their concentrations diminish and when the antibody and antigen reach _____ , they form a _____.

The _____ is formed by the lattice structure (_____) formed by the _____ and _____ complexes.
Double immunodiffusion gel precipitation analyses are unique in that they allow for determination of antigenic relatedness of an unknown test material with a known antigen. Specifically allows the determination of whether the unknown is identical, partially identical, or nonidentical with the known antigen.

Double diffusion gel precipitation can be used quantitatively to determine relative concentrations of the antigens based on where they form a precipitin line. The antigen with a higher concentration would form a precipitin line closer to the antibody well because it would diffuse further before it decreased to the level needed for equivalence.
_____ analyses are unique in that they allow for determination of _____ relatedness of an unknown test material with a known _____. Specifically allows the _____ of whether the unknown is _____, _____ or _____ with the known antigen.

_____ can be used _____ to determine relative concentrations of the _____ based on where they form a _____. The antigen with a _____ would form a precipitin line closer to the _____ well because it would diffuse further before it _____ to the level needed for _____ .
Double immunodiffusion can be used to compare antigenic relatedness. Two different antigens are placed in different wells and the antiserum is placed in a third. One of three patterns can occur:

Identity - precipitin line forms an arch
Nonidentity - precipitin lines form a cross
Partial identity - precipitin line forms a spur.
_____ can be used to compare antigenic _____. Two different _____ are placed in different wells and the _____ is placed in a third. One of _____ patterns can occur:

_____ - precipitin line forms a _____
_____ - precipitin line forms a _____
_____ - precipitin line forms a _____
Radial immunodiffusion gel contains the antibody evenly distributed throughout.

A standard curve is prepared by using three different concentrations of the antigen in three different wells using the diameter of the precipitin line.

Approximate concentration of an antigen can be obtained by comparing the diameter of the precipitin line to the standard curve using two different methods, the Fahey method and Mancini method.
_____ contains the _____ evenly distributed throughout.

A _____ is prepared by using _____ different concentrations of the _____ in _____ different wells using the diameter of the _____.

_____ concentration of an antigen can be obtained by comparing the _____ of the _____ to the standard curve using two different methods, the _____ method and _____ method.
Radial immunodiffusion methods
Fahey method (Kinetic method) -
The diffusion proceeds for 18 hours and the diameter is proportional to the log of the concentration. It is plotted on semi-log paper using the diameter of the precipitin line on the arithmetic axis and concentration on the y-axis.

Mancini method - (Endpoint method)
The diffusion is allowed to come to equilibrium in 48 to 72 hours. The square of the diameter is directly proportional to the concentration.
Radial immunodiffusion methods
_____ (_____ method) -
The diffusion proceeds for _____ hours and the diameter is _____ to the _____ of the concentration. It is plotted on _____ paper using the diameter of the precipitin line on the _____ axis and concentration on the _____.

_____ (_____ method) -
The _____ is allowed to come to equilibrium in _____ hours. The _____ of the diameter is directly _____ to the concentration.
Radial immunodiffusion is used to measure IgG, IgM, and IgA levels. Serum IgE and IgD are too low to be reliably measured by this method. The antibodies are being measured as antigens while the antibody being used is anti-human immunoglobulin that is Fc specific. Concentrations of complement are determined this way as well.
_____ is used to measure _____, _____, and _____ levels. Serum _____ and _____ are too low to be reliably measured by this method. The _____ are being measured as antigens while the antibody being used is _____ that is Fc specific. Concentrations of _____ are determined this way as well.
The discovery that serum could agglutinate bacteria happened just before the 20th century. This contributed evidence that the humoral part of the blood help provide defense against infection. This was soon used to diagnose typhoid fever.

Agglutination is not very sensitive because nothing amplifies the actual antibody-antigen reaction, however it is more sensitive than precipitation because the larger antigen particles enhance visualization. The increased sensitivity of agglutination of precipitation was the reason for the development of passive agglutination.
The discovery that serum could _____ bacteria happened just before the _____ century. This contributed _____ that the _____ part of the blood help provide defense against _____. This was soon used to diagnose _____.

_____ is not very sensitive because nothing _____ the actual _____ reaction, however it is more _____ than _____ because the larger antigen particles enhance _____. The increased _____ of agglutination over precipitation was the reason for the development of _____.
Types of agglutination:
direct
passive
reverse passive
inhibition - serum is inoculated with antigen, uses latex beads covered with an antibody looking for an antigen
Types of agglutination:
_____
_____
_____
_____ - serum is _____ with antigen, uses _____ covered with an _____ looking for an _____
Direct agglutination naturally have antigen on their surface and tests for the presence of an antibody and a positive test result is one causes agglutination.
Direct agglutination naturally have _____ on their surface and tests for the presence of an _____ and a positive test result is one _____ agglutination.
Passive agglutination uses latex beads that are coated with an antigen and tests for the presence of an antibody and a positive test result is one causes agglutination.
Passive agglutination uses _____ that are coated with an _____ and tests for the presence of an _____ and a positive test result is one _____ agglutination.
Reverse passive agglutination uses latex beads that are coated with an antibody and tests for the presence of an antigen and a positive test result is one that cause agglutination.
Reverse passive agglutination uses latex beads that are coated with an _____ and tests for the presence of an _____ and a positive test result is one that _____ agglutination.
Agglutination inhibition reaction uses serum that is inoculated with and antigen. Latex bead that is coated with antigen an tests for the presence of an antigen and a positive test result is one that does not cause agglutination.
Agglutination _____ reaction uses serum that is inoculated with an _____ . Latex bead that is coated with _____ and tests for the presence of an _____ and a positive test result is one that _____ agglutination.
Rosalyn Yalow, in 1959, developed the first assay, radioimmunoassay (RIA), in which marker or label was covalently attached to the antibody or antigen to increase the sensitivity of the detection for which she won the Nobel prize.

Yalow's assay which was developed to help study the clearance of insulin, is the predecessor to many sensitive assays using detectable markers or labels to increase sensitivity with a thousandsfold increase. Yalow compared this to viewing the sky with a powerful telescope versus the eye alone.

Engvall and Pelman, in the 1970s, developed an enzyme label for antigen-antibody reactions to use instead of a radioactive tag.
_____, in _____, developed the first assay, _____ (_____), in which marker or label was _____ attached to the antibody or antigen to increase the _____ of the detection for which she won the _____.

_____ assay which was developed to help study the clearance of _____, is the predecessor to many sensitive assays using detectable _____ or _____ to increase _____ with a thousandsfold increase. _____ compared this to viewing the _____ with a powerful telescope versus the _____ alone.

_____ and _____ , in the _____ , developed an _____ label for antigen-antibody reactions to use instead of a _____ tag.
Not only can radiolabels be used, but antibody or antigen can be alternatively labeled with:
enzymes (enzyme immunoassays (EIA) or enzyme-linked immunosorbant assays (ELISA))
fluorescent compounds (fluorescent immunoassay (FIA)
chemiluminescent labels
colloid particles
Not only can radiolabels be used, but antibody or antigen can be alternatively labeled with:
_____
_____
_____
_____
In labeled immunoassays reactions, its essential that the label can be easy to measure and to separate the bound label from the unbound labels amounts. This is usually done by using a solid phase for the reaction and washing away the unbound reactant.
In _____ immunoassays reactions, its essential that the _____ can be easy to _____ and to _____ the _____ label from the _____ labels amounts. This is usually done by using a _____ for the reaction and _____ away the unbound reactant.
Heterogeneous assays require separating the bound labels from the free labels, while homogeneous assays do not require this step. Homogeneous assays can be accomplished because the binding of an antigen to an antibody affects the activity of the label in such a way as to cause a measurable change.
_____ assays require _____ the bound labels from the free labels, while _____ assays do not require this step. _____ assays can be accomplished because the binding of an _____ to an _____ affects the _____ of the label in such a way as to cause a _____ change.
Radiolabels can be detected by gamma counters or liquid scintillation counters.

Enzyme labels can cause a production of color, fluorescence, flash of light, or change in optical reflection. They are measured using a spectrophotometer, fluorimeter, or luminometer.

Fluorescent labels are detected in a spectrofluorimeter by detecting the amount of fluorescence at the emission wavelength after application of the excitation wavelength of light.

Chemiluminescent labels are measured by a luminometer once the compound is activated with a "trigger".
Radiolabels can be detected by _____ counters or _____ counters.

Enzyme labels can cause a production of _____, _____, _____, or change in _____. They are measured using a _____, _____, or _____.

Fluorescent labels are detected in a _____ by detecting the amount of fluorescence at the _____ after application of the _____ of light.

Chemiluminescent labels are measured by a _____ once the compound is activated with a "_____ ".
Types of heterogeneous assays:
direct immunoassay
indirect immunoassay
capture/sandwich immunoassay
competitive assay
Types of heterogeneous assays:
_____
_____
_____
_____
Most heterogeneous assays are performed using solid-phase attachment of one of the reactants to a microtiter plate, a slide, or a bead. The use of the solid phase allows the rapid separation of the bound molecules from the free ones by washing steps to remove free labeled reagent so that only the bound label is measured.
Most _____ assays are performed using _____ attachment of one of the reactants to a _____, _____, or a _____. The use of the solid phase allows the _____ of the bound molecules from the free ones by _____ steps to remove free labeled reagent so that only the bound label is _____.
Direct immunoassays utilize a labeled antibody binding to an antigen or labeled antigen binding to an antibody.

It is typically used to detect an antigen in a cell preparation or biopsy sample.

An example is a direct fluorescent immunoassay to detect anthrax antigen in a suspected skin lesion biopsy.
_____ immunoassays utilize a _____ antibody binding to an _____ or _____ antigen binding to an _____.

It is typically used to detect an antigen in a _____ or _____.

An example is a _____ to detect _____ antigen in a suspected _____.
Indirect immunoassays utilize an unlabeled antigen, an unlabeled antibody, and a labeled antiglobulin to detect the reaction of the initial antibody and antigen complex.

This is typically used to measure a patient's antibody titer to a known antigen.

An example is an indirect enzyme immunoassay to measure a patient's titer or level of antibody to rubella.

Labeled protein A, isolated from Staphylococcus aureus, can be used instead of a labeled anti-human IgG antibody.
_____ immunoassays utilize an _____ antigen, an _____ antibody, and a _____ antiglobulin to detect the reaction of the initial _____ and _____ complex.

This is typically used to measure a patient's _____ to a known antigen.

An example is an _____ to measure a patient's _____ or level of antibody to _____.

Labeled _____, isolated from _____, can be used instead of a labeled _____ antibody.
Sandwich assays usually capture an antigen between 2 molecules of antibody, one of which captures the antigen to a solid phase while the other is labeled and used to visualize the reaction.

The assay is commonly performed on disposable membrane cassettes.

An example would be a pregnancy colloid labeled immunochromatographic sandwich assay.

An antigen must have more than one epitope to utilize a sandwich assay.
_____ assays usually _____ an antigen between 2 molecules of _____, one of which captures the _____ to a solid phase while the other is _____ and used to _____ the reaction.

The assay is commonly performed on disposable _____.

An example would be a _____ sandwich assay.

An _____ must have more than one _____ to utilize a sandwich assay.
Sandwich assays:
Adaptions have been developed to ensure that an assay for antibody is IgM specific because IgG positivity may indicate only a past infection and since IgM is only around for the first few weeks of an infection and thus indicates an acute infection with this pathogen.

IgM determinations can be negatively affected by the presence of IgG because IgG has a higher affinity and will preferentially bind to the antigen over IgM giving a false negative to the presence of IgM to the antigen.

False positive for IgM can occur if the patient has IgG to a pathogen and has IgM rheumatoid factor, but adaptations have been developed to detect pathogen-specific IgM without interference with serum IgG to the pathogen.
Sandwich assays:
_____ have been developed to ensure that an assay for antibody is _____ specific because _____ positivity may indicate only a _____ infection and since _____ is only around for the first few weeks of an infection and thus indicates an _____ with this pathogen.

_____ determinations can be _____ affected by the presence of _____ because _____ has a higher affinity and will preferentially _____ to the antigen over _____ giving a false _____ to the presence of _____ to the antigen.

_____ for _____ can occur if the patient has _____ to a pathogen and has _____ rheumatoid factor, but adaptations have been _____ to detect pathogen-specific _____ without interference with serum _____ to the pathogen.
Competitive assays are used when an antigen is small and has either only one epitope or if binding to multiple epitopes would create steric hindrance.

Competitive immunoassay uses a limited reagent that competes with the analyte in the patient's sample.

Antigen can compete for reaction with the antibody, or the antibody can compete for the antigen.

Another example is the a microparticle enzyme immunoassay for antibody to hepatitis A antigens. The amount of patient antibody is inversely related to the amount of substrate conversion.
_____ assays are used when an antigen is _____ and has either only one _____ or if binding to multiple _____ would create _____.

_____ immunoassay uses a limited _____ that competes with the _____ in the patient's sample.

_____ can compete for reaction with the antibody, or the _____ can compete for the antigen.

Another example is the a _____ enzyme immunoassay for antibody to _____ antigens. The amount of patient antibody is _____ related to the amount of substrate _____.
Heterogeneous assays can be performed utilizing any type of labels. The choice of radioactive, enzyme, fluorescent, chemiluminescent, or colloid particle labels is based on many factors, including the equipment available, cost, ease of use, sensitivity, and personal preference.
Heterogeneous assays can be performed utilizing _____ type of labels. The choice of _____, _____, _____, _____, or _____ labels is based on many factors, including the _____, _____, _____, _____, and _____.
Radioimmunoassays were the first labeled immunoassays developed and thought to be ideal because they caused no steric effects. It was later determined that enzymatic labels also had little to no steric effects.

A disadvantage of radio labels is that they have a half=life of 59.6 days, but they are highly sensitive.
_____were the first labeled immunoassays developed and thought to be _____because they caused no ____. It was later determined that _____labels also had little to no ____.

A disadvantage of _____is that they have a half-life of _____days, but they are highly ____.
Enzyme labels are the must often used for immunoassays. The most frequently used are horseradish peroxidase and alkaline phosphatase. Less common enzymes include glucose oxidase, B-galactosidase, and glucose-6-phosphate dehydrogenase.
_____ labels are the must often used for immunoassays. The most frequently used are _____ and ____. _____ enzymes include glucose oxidase, B-galactosidase, and glucose-6-phosphate dehydrogenase.
Horseradish Peroxidase is sensitive to a number of inhibitors, so all buffers and solutions must be free of azides, sulfides, and cyanides. Substrate solutions are light sensitive, and many must be made just prior to use.
_____ is sensitive to a number of _____ , so all buffers and solutions must be free of _____, _____, and _____. Substrate solutions are _____ and many must be made just prior to use.
Phosphates should be avoided in buffers from the alkaline phosphatase subsrtate and conjugate because their use can lower the assay's sensitivity. Cysteine, cyanides, arsenates, and divalent cation chelators can also impact the sensitivity of the alkaline phosphatase-labeled assay.
Phosphates should be avoided in buffers from the _____ subsrtate and _____ because their use can lower the assay's sensitivity. _____, _____, _____, and _____ chelators can also impact the _____ of the _____-labeled assay.
Types of homogeneous assays:
fluorescence polarization immunoassay
enzyme immunoassay
Types of homogeneous assays:
_____
_____
In the fluorescence polarization immunoassay, a fluorescent label is placed on the small molecule that is the analyte.

Small molecules rotate freely in solution, so when plane polarized light is used as an excitation wavelength for a fluorochrome attached to a small molecule, this free rotation results in the emission of the fluorescent light in a variety of directions so that the light is no longer in 1 plane.

If a relatively large antibody molecule has bound this small molecule, the rotation is retarded, and when the fluorochrome is excited, the emission remains polarized.

This is a competitive immunoassay and is used to test for drugs.

Positive result will have less polarization and negative will have more polarization.
In the _____ immunoassay, a _____ label is placed on the small molecule that is the _____.

Small molecules _____ in solution, so when _____ light is used as an excitation wavelength for a _____ attached to a small molecule, this _____ results in the emission of the _____ in a variety of directions so that the light is no longer in _____.

If a relatively _____ molecule has bound this small molecule, the rotation is _____, and when the _____ is excited, the emission remains _____.

This is a _____ immunoassay and is used to test for _____.

Positive result will have _____ polarization and negative will have _____ polarization.
In a homogeneous enzyme immunoassay, an enzyme is bound to the drug analyte can perform its enzymatic function when the drug is free in solution. However, when an antibody binds to the drug-enzyme complex, the enzyme is sterically inhibited from having its enzymatic effect.

This is a competitive immunoassay and is used to test for methamphetamine.

Positive result will have more enzymatic activity than a negative result.
In a _____ immunoassay, an _____ is bound to the drug _____ can perform its _____ function when the drug is free in _____. However, when an _____ binds to the drug-enzyme complex, the enzyme is _____ from having its _____ effect.

This is a _____ immunoassay and is used to test for _____.

Positive result will have _____ enzymatic activity than a negative result.
Antibody Detection (Heterogeneous Assays)
Noncompetitive - strength of color is proportional to the concentration of patient antibody
Competitive - strength of the color is inversely proportional to the concentration of the patient antibody
Capture - strength of color is proportional to the concentration of patient antibody
Antibody Detection (Heterogeneous Assays)
_____ - strength of color is _____ to the concentration of patient antibody
_____ - strength of color is _____ to the concentration of the patient antibody
_____ - strength of color is _____ to the concentration of patient antibody
A depiction of the amount of precipitin that would form with different rations of antigen and antibody.

The prozone has antibody excess, and only small complexes with 2 antibody molecules and 1 antigen form.

At equivalence, the number of paratopes roughly equals the number of epitopes, and the larges complexes form.

The postzone has excess antigen, and the small complexes are formed of 2 antigens per antibody molecule.
Ouchterlony double diffusion gel precipitation assay

Identity
Ouchterlony double diffusion gel precipitation assay

Non-identity
Ouchterlony double diffusion gel precipitation assay

Partial identity
Radial immunodiffusion
Turbidometry
Nephalometry
Summary of Precipitation Reactions
Summary of Agglutination Reactions
Direct Immunoassays
Indirect Immunoassay
Sandwich / Capture Immunoassay
Competitive Assay
Steps of the Heterogeneous Immunoassays
Flourescence polarization immunoassay
Homogeneous enzyme immunoassay
The Central Dogma Theory was first coined by Francis Crick in 1953. It can only flow in one direction in humans an involves DNA, RNA, and proteins. It has three processes called replication, transcription, and translation.
The _____ was first coined by _____ in _____. It can only flow in one direction in _____ an involves _____, _____, and _____. It has three processes called _____, _____, and _____.
DNA was discovered in 1869 by Johann Friedrich Miescher, by isolating a viscous substance from the nuclei of white blood cells which he called nuclein.

Oswald Avery proved DNA was the carrier of genetic information in 1943.

James Watson and Francis Crick discovered the structure of DNA in 1953.
DNA was discovered in _____ by _____, by isolating a _____ substance from the nuclei of _____ which he called _____ .

_____ proved DNA was the carrier of genetic information in _____.

_____ and _____ discovered the structure of DNA in _____.
Deoxyribonucleic acid is found in a supercoiled double helix confirmation. It is made up of paired nucleotides which are formed by phosphorylated ribose sugar and a nitrogen base.

The purines are guanine and adenine and have a single ring structure while the pyrimidines are cytosine and thymine and have a double ring structure.

Adenine pairs with thymine while guanine pairs with cytosine.
_____ is found in a _____ double helix confirmation. It is made up of paired _____ which are formed by _____ and a _____.

The purines are _____ and _____ and have a _____ ring structure while the pyrimidines are _____ and _____ and have a _____ ring structure.

Adenine pairs with _____ while guanine pairs with _____.
Replication of DNA takes place in the nucleus and it done semi-conservatively. Each molecule consists of one template strand and one newly synthesized strand.
Replication of DNA takes place in the _____ and it done _____. Each molecule consists of one _____ and one newly _____ strand.
Transcription is the process of transcribing DNA into mRNA so that it may be taken to the ribosome to be translated into protein and takes place in the nucleus.
Transcription is the process of _____ DNA into _____ so that it may be taken to the _____ to be _____ into _____ and takes place in the _____.
Translation is the process in which the genetic code, or nucleic acid sequence, is translated to amino acid sequence and, ultimately, to phenotype. This process takes place in the ribosomes.
Translation is the process in which the _____, or nucleic acid sequence, is translated to _____ and, ultimately, to _____. This process takes place in the _____.
Ribonucleic acid is produced as a single strand which can fold in upon itself to take on a double stranded character that is important to its function. It can also bind to complementary DNA or RNA to form a double helix. RNA does not contain thymine but uracil instead.
_____ is produced as a _____ strand which can _____ in upon itself to take on a _____ character that is important to its _____. It can also bind to complementary _____ or _____ to form a _____ helix. RNA does not contain _____ but _____ instead.
Types of RNA:
ribosomal (rRNA) - 80-90% of cellular RNA
messenger (mRNA) - the link between DNA and proteins
transfer (tRNA) - transports amino acids to the ribosome to produce proteins. Can recognize both nucleic acid and protein sequence components.
Types of RNA:
_____ - 80-90% of cellular _____
_____ - the _____ between DNA and _____
_____ - _____ amino acids to the _____ to produce proteins. Can recognize both _____ and protein sequence components.
Proteins are the ultimate products of transcription and translation and are the organism's phenotype. They are composed of amino acid polymers. Each amino acid contributes to the shape and function of the proteins and a single protein can be made up of several domains with different properties.
_____ are the ultimate products of transcription and _____ and are the organism's _____. They are composed of _____. Each amino acid contributes to the _____ and _____ of the proteins and a single protein can be made up of several _____ with different _____.