Block 5 Flashcards by catherine perea

CAPTURE ANTIBODY GRABS THE
ANTIGEN
ENZYME LABELLED ANTIBODY THEN
ATTACHES TO THE ANTIGEN
ANTIBODY-ANTIGEN-ANTIBODY
SANDWICH

this is called….

SANDWICH ELISA

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what do you use to diagnose an unknown antigen, food product quality control or for allergy tesing?

SANDWICH ELISA

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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

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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

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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

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SEROLOGICAL ASSAYS FOR DETECTION

FLUORESCENCE ANTIBODY TEST (FAT)

with an unknown antigen do you use a direct or indirect FAT?

direct

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* 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

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FLUORESCENCE ANTIBODY TEST (FAT)

with unknown antibody -
direct or indirect FAT?

INDIRECT FAT

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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

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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

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* FLUORESCENCE ANTIBODY TEST (FAT)

This one is pretty good:

Direct is a shorter word than indirect—>
Antigen is a shorter word than….

antibody

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IMMUNOHISTOCHEMISTRY

can it be both indirect and direct?

CAN BE DIRECT OR INDIRECT

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IMMUNOHISTOCHEMISTRY

THE ANTIBODY IS TAGGED WITH AN ENZYME, GENERALLY
?
WHICH BINDS TO A TARGET
ANTIGEN.

HORSERADISH PEROXIDASE

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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

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IMMUNOHISTOCHEMISTRY

this is an example or indirect or direct?

indirect

*just checking to see if you were REALLY looking at the picures…

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IMMUNOHISTOCHEMISTRY

this is an example of indirect or direct?

direct

*just checking to see if you were REALLY looking at the picures…

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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

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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

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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.

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SEROLOGICAL ASSAYS

this is an example of what test?

AGAR GEL

IMMUNODIFFUSION TEST

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SEROLOGICAL ASSAYS

COMPLEMENT FIXATION TEST

or + RESULTS WILL LEAD TO LYSIS OF
RBCS AND WILL HAVE NO ANTIBODIES IN
THE SERUM OF THE PATIENT.

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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.

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POSITIVE RESULTS

NEUTRALIZATION ASSAYS

NEUTRALIZATION OF A VIRUS IS DEFINED AS THE LOSS OF
INFECTIVITY THROUGH REACTION OF THE VIRUS WITH ?

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SPECIFIC ANTIBODY

Virus + Ab=inactive virus

POLYMERASE CHAIN REACTION (PCR)

what is it?

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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.

Block 5 Flashcards

(50 cards)