Block 5 Flashcards
- CAPTURE ANTIBODY GRABS THE
ANTIGEN - ENZYME LABELLED ANTIBODY THEN
ATTACHES TO THE ANTIGEN - ANTIBODY-ANTIGEN-ANTIBODY
SANDWICH
this is called….
SANDWICH ELISA
what do you use to diagnose an unknown antigen, food product quality control or for allergy tesing?
SANDWICH ELISA
how does a sandwich elisa detect allergies or alergen proteins?
The technique is used for allergy testing. The test
will capture antibodies from blood samples
towards specific foods or other typical allergens.
ELISA can also be used to detect allergen proteins
in food products for quality control
TYPES OF ELISA CONTINUED
A DECREASE IN SIGNAL WHEN COMPARED TO ASSAY
WELLS WITH PURIFIED AG ALONE WHICH INDICATED THE PRESENCE OF AG IN
THE SAMPLE. WEAK SIGNAL EQUALS POSITIVE RESULT
this is called?
COMPETITIVE ELISA
COMPETITIVE ELISA
1* ANTIGEN AND ANTIBODIES ARE INCUBATED TOGETHER SO THEY FORM
A COMPLEX
2* WASH OUT THE UNBOUND ANTIBODY
What is step 3?
4* ADD THE SUBSTRATE
5* THE SECONDARY ANTIBODY BOUND WITH THE PRIMARY ANTIBODY
WILL CHANGE COLOR
3* ENZYME TAGGED SECONDARY LABELED ANTIBODY ADDED AND BINDS
TO THE PRIMARY ANTIBODY
SEROLOGICAL ASSAYS FOR DETECTION
FLUORESCENCE ANTIBODY TEST (FAT)
with an unknown antigen do you use a direct or indirect FAT?
direct
* FLUORESCENCE ANTIBODY TEST (FAT)
IFAT EMPLOYS A SECONDARY
ANTIBODY LABELED WITH A FLUORESCENT
MARKER THAT RECOGNIZES THE PRIMARY
ANTIBODY BOUND TO THE ANTIGEN
is this direct or indirect FAT?
INDIRECT FAT
FLUORESCENCE ANTIBODY TEST (FAT)
with unknown antibody -
direct or indirect FAT?
INDIRECT FAT
FLUORESCENCE ANTIBODY TEST (FAT)
LABELLED ANTIBODIES ARE ADDED
ONTO THE SAMPLE (ANTIGEN) LEADING TO
FLUORESCENCE AT THE BINDING SITE OF SPECIFIC
ANTIBODIES.
is this direct or indirect FAT
DIRECT FAT
FLUORESCENCE ANTIBODY TEST (FAT)
LABELLED ANTIBODIES ARE ADDED
ONTO THE SAMPLE (ANTIGEN) LEADING TO
FLUORESCENCE AT THE BINDING SITE OF SPECIFIC
ANTIBODIES.
is this direct or indirect FAT
DIRECT FAT
* FLUORESCENCE ANTIBODY TEST (FAT)
This one is pretty good:
Direct is a shorter word than indirect—>
Antigen is a shorter word than….
antibody
IMMUNOHISTOCHEMISTRY
can it be both indirect and direct?
CAN BE DIRECT OR INDIRECT
IMMUNOHISTOCHEMISTRY
- THE ANTIBODY IS TAGGED WITH AN ENZYME, GENERALLY
?
WHICH BINDS TO A TARGET
ANTIGEN.
HORSERADISH PEROXIDASE
IMMUNOHISTOCHEMISTRY
THE ENZYME
does what?
THAT CAN BE VISUALIZED IN THE
INFECTED CELLS WITH A STANDARD LIGHT MICROSCOPE.
REACTS WITH A SUBSTRATE TO PRODUCE A
COLORED PRODUCT
IMMUNOHISTOCHEMISTRY
this is an example or indirect or direct?
indirect
*just checking to see if you were REALLY looking at the picures…
IMMUNOHISTOCHEMISTRY
this is an example of indirect or direct?
direct
*just checking to see if you were REALLY looking at the picures…
IMMUNOCHROMATOGRAPHY
(LATERAL FLOW DEVICES)
A FORM OF
?
THAT IS
SIMPLE TO PERFORM, EASY TO CARRY, AND DOES
NOT REQUIRE SPECIALIZED EQUIPMENT. FOR
EXAMPLE: ?
** POC (POINT OF CARE) TEST**
A PREGNANCY TEST
AGGLUTINATION TESTS
SPECIFIC
?
(EITHER ON THE PATHOGEN OR LATEX BEADS) CREATING
SINGULAR CLUMPS AND FORMING LARGER COMPLEXES
THAT ARE EASILY PRECIPITATED.
ANTIBODIES ARE BOUND TO ANTIGENS
HEMAGGLUTINATION AND
HEMAGGLUTINATION
INHIBITION TEST
THE HEMAGGLUTINATION AND THE INHIBITION
METHOD RELIES ON THE PROPERTY OF SOME …
what to do what?
PATHOGENS TO NONSPECIFICALLY AGGLUTINATE
ERYTHROCYTES.
SEROLOGICAL ASSAYS
this is an example of what test?
AGAR GEL
IMMUNODIFFUSION TEST
SEROLOGICAL ASSAYS
COMPLEMENT FIXATION TEST
- or + RESULTS WILL LEAD TO LYSIS OF
RBCS AND WILL HAVE NO ANTIBODIES IN
THE SERUM OF THE PATIENT.
NEGATIVE RESULTS
SEROLOGICAL ASSAYS
COMPLEMENT FIXATION TEST
- or + RESULTS WILL LEAD TO A RBC
PELLET AT THE BOTTOM AND WILL HAVE
ANTIBODIES IN THE PATIENT’S SERUM.
POSITIVE RESULTS
NEUTRALIZATION ASSAYS
NEUTRALIZATION OF A VIRUS IS DEFINED AS THE LOSS OF
INFECTIVITY THROUGH REACTION OF THE VIRUS WITH ?
SPECIFIC ANTIBODY
Virus + Ab=inactive virus
POLYMERASE CHAIN REACTION (PCR)
what is it?
- AMPLIFICATION OF VIRAL GENOME/DNA
POLYMERASE CHAIN REACTION (PCR)
- HOW IT WORKS?
- DENATURATION
- ANNEALING
- EXTENSION/ELONGATION
POLYMERASE CHAIN REACTION (PCR)
REAL TIME/QUANTITATIVE PCR: ALLOWS for what?
ALLOWS
MONITORING AND QUANTIFICATION OF
INCREASING ACCUMULATION OF PCR
PRODUCTS/NUCLEIC ACID LOAD AS
REACTION OCCURS.
POLYMERASE CHAIN REACTION (PCR)
is very useful for the study of what in the patient?
VERY USEFUL TO STUDY VIRUS LOAD IN THE
PATIENT
REFERS TO THE PROCESS BY WHICH THE SEQUENCE OF
BASES IN A DNA MOLECULE IS ELUCIDATED AND READ.
GENOME SEQUENCING DNA SEQUENCING
GENOME SEQUENCING
WHY IT’S IMPORTANT?
there are 7 reasons
WHY IT’S IMPORTANT?
* 1. PATHOGEN DETECTION
* 2. STUDIES ON GENETIC VARIATION, SUCH AS GENOTYPING,
EVOLUTION AND INTERSPECIES TRANSMISSION OF PATHOGENS
* 3. IDENTIFICATION OF NOVEL AND UNDISCOVERED STRAINS
* 4. DEVELOPMENT OF DIAGNOSTICS, SUCH AS GENOTYPING PRIMERS
OR PROBES
* 5. IDENTIFICATION OF GENES ASSOCIATED WITH DRUG RESISTANCE
* 6. DEVELOPMENT OF THERAPEUTICS
* 7. JUDGING THE EFFICACY OF CURRENT VACCINES AND FORMULATING
NEW VACCINE STRATEGIES
GENOME SEQUENCING
this describes what?
SIGNIFICANTLY CHEAPER, QUICKER, NEEDS
SIGNIFICANTLY LESS DNA, HAS HIGH
THROUGHPUT, AND IS MORE ACCURATE AND
RELIABLE THAN SANGER SEQUENCING.
NEXT GENERATION
SEQUENCING
METAGENOMICS
THE STUDY OF
?
THE IN A SAMPLE BY ANALYZING THE
SAMPLE’S
?
CONTENT, AND IS A POWERFUL METHOD FOR
RANDOM DETECTION OF EXISTING AND NEW
PATHOGENS.
COLLECTIVE SET OF MICROBIAL
POPULATIONS
ENTIRE NUCLEOTIDE SEQUENCE
THE USE OF VIRUS GENOME SEQUENCE DATA TO
STUDY EVOLUTION OF VIRUSES AND GENETIC RELATIONSHIPS AMONG VIRUSES.
EXAMPLE: PORCINE ROTAVIRUS IS GENETICALLY CLOSE TO HUMAN ROTAVIRUS.
this is called what?
PHYLOGENETIC ANALYSIS
MICROARRAYS:
- USES SEVERAL THOUSANDS OF KNOWN DNAS (PROBES), AMPLIFIED BY
PCRS/RT-PCRS ARE SPOTTED ONTO A GLASS OR SILICON CHIP. - TARGET/SAMPLE DNA ARE ?
- POSITIVE REACTION = ?
- ADVANTAGE: ?
FLUORESCENTLY LABELLED AND
HYBRIDIZED/ADDED TO CHIP. WITH THE DNA PROBES
FLUORESCENT SIGNAL WHERE PROBE DNA IS IN CHIP.
SURVEILLANCE STUDIES OF HUNDREDS OF PATHOGENS CAN BE
DONE SIMULTANEOUSLY WITH SINGLE CHIP.
The study of ciris evolution using genome sequence data is known as?
a-metagenomics
b-phylogenetics
c-PCP
d-microarray
b-phylogenetics
TREATMENT OF VIRAL DISEASES
ANTIVIRAL DRUGS: INTERFERE WITH ????
THESE ARE A CLASS OF MEDICATION USED SPECIFICALLY
TO TREAT VIRAL INFECTIONS.
VIRAL REPLICATION.
TREATMENT OF VIRAL DISEASES
IMMUNE SYSTEM STIMULATION: ??? HAVE
ANTIVIRAL EFFECTS AND MODULATE THE FUNCTIONS OF
THE IMMUNE SYSTEM.
INTERFERONS
TREATMENT OF VIRAL DISEASES
SYNTHESIZE ????
–>
passive ????
ANTIBODIES
immunity
Aciclovir is converted by what process? to Aciclovir monophosphate, which is then converted by host cell kinases to ? which is a very potent inhibitor of viral DNA replication and competitively inhibits and inactivates the viral DNA polymerase. The monosphosphate form also incorporates ito the viral DMA, resulting in what?
viral thymidine kinase
aciclovir triphosphate (ACV-TP also known as aciclo-GTP
chain termination
ACYCLOVIR (HERPES VIRUS)
ADMINISTERED IN THE ?? FORM AND REQUIRES ??
TO CONVERT IT INTO THE FORM WHICH MAY INTERFERE WITH
VIRUS REPLICATION.
INACTIVE
VIRAL ENZYMES
ACTIVE
ACYCLOVIR (HERPES VIRUS)
IS IT A NON-SYNTHETIC NUCLEOSIDE ANALOG OF DEOCYGUANOSINE?
NO
SYNTHETIC
ACYCLOVIR (HERPES VIRUS)
IS NON-TOXIC TO THE UNINFECTED HOST CELL BECAUSE ??
THEY CANNOT
BE PHOSPHORYLATED AND INCORPORATED INTO THE HOST DNA
ACYCLOVIR (HERPES VIRUS)
IT IS USED FOR THE TREATMENT OF WHAT 3 THINGS?
- TREATMENT OF:
- HERPESVIRUS INFECTIONS IN HUMANS
- FELINE HERPESVIRUS 1 INDUCED CORNEAL ULCERS
- EQUINE HERPESVIRUS-1 INDUCED ENCEPHALOMYELITIS
MECHANISM OF ANTIVIRAL EFFECT OF ACYCLOVIR
3 STEPS IN THE FIRST STEP:
Phasphates are cleaved from ? to form acyclovirmonophosphate by the herpes virus DNA polymerase
acyclovir-triphosphate
MECHANISM OF ANTIVIRAL EFFECT OF ACYCLOVIR
3 STEPS IN THE SECOND STEP:This DNA polymerase then incorporates the acyclovir monophosphate into the
growing DNA strand causing viral DNA chain synthesis to ???
stop elongation.
MECHANISM OF ANTIVIRAL EFFECT OF ACYCLOVIR
3 STEPS IN THE THIRD STEP: Competitive inhibition of viral DNA polymerase: HOW?
The acyclovir triphosphates
compete with dGTPs (Guanine nucleotide) for viral DNA polymerase.
??? INHIBITS REPLICATION OF MOST
STRAINS OF INFLUENZA A VIRUSES BY BLOCKING
UNCOATING OF THE VIRUS.
AMANTADINE (INFLUENZA A)
AMANTADINE (INFLUENZA A)
- MECHANISM:
- THE M2 ION CHANNEL IS THE TARGET VIA CLOGGING
AND PREVENTING PUMPING OF PROTONS INTO
VIRIONS. - THEREFORE , WHAT?
- THEREFORE , THE VIRAL RNA’S REMAIN BOUND TO THE
M1 AND CANNOT ENTER THE NUCLEUS INHIBITING
REPLICATION.
NUCLEOSIDE ANALOG REVERSE
TRANSCRIPTASE INHIBITORS (NRTIS)
***THINK AIDS WITH AZT
ZIDOVUDINE OR AZIDOTHYMIDINE (ZVD/AZT):
RESEMBLE THE DEOXYRIBONUCLEOTIDE THAT CONTAINS ???
INSERTION OF MONOPHOSPHATE INTO CDNA DOES WHAT?
AZT IS SHOWN TO REDUCE CLINICAL SIGNS OF ???
THYMINE
BLOCKS
GROWTH/TRANSCRIPTION
FIV-
POSITIVE CATS WHEN GIVEN 10MG/KG BID, SQ FOR 3
WEEKS.
IMPORTANT
THINGS TO
REMEMBER
??? ARE REQUIRED TO CLEAVE THE HIV
POLYPROTEINS INTO FUNCTIONAL PROTEINS.
PROTEASES AKA PEPTIDASE, ENZYME THAT CATALYZES (SPEEDS UP) THE BREAKDOWN OF PROTEINS
IMPORTANT
THINGS TO
REMEMBER
PROTEASE INHIBITORS EXIST AND CAN DO WHAT RE: hiv?
PREVENT hiv FROM CLEAVING AND MATURING AND NON-INFECTIOUS VIRUSES PRODUCE.
