Laboratory Diagnosis

Question 1

2 types of examination

Answer 1

Direct and Indirect Examination

Question 2

Specimen is what?

Answer 2

Examined Directly

Question 3

what can be directly examined under a specimen

Answer 3

• Virus Particles
• Viral Antigen
• Inclusion Bodies
• Virus Nucleic Acid

Question 4

what are the examination under direct examination

clue; ACCEN (asin)

Answer 4

• Antigen Detection
• Electron Microscopy
• Cytology (Light Microscopy)
• Classical Molecular Technique
• Newer Molecular Techniques

Question 5

what are the test found under Antigen Detection

Answer 5

1. Immunofluoresence (IF)
2. ELISA (Enzyme-Linked Immunosorbent Assay)
3. Latex Agglutination

Question 6

Immunofluoresence (IF)

dye used

Answer 6

Dye Fluorescein Isothiocyanate

Question 7

Immunofluoresence (IF)

Positive result

Answer 7

Apple Green Florescence with reddish brown counterstained cells

Question 8

Immunofluoresence (IF)

Result

Answer 8

Viral antigen is detected or Viral antigen is not detected

Question 9

what should be done with Immunofluoresence result?

Answer 9

• Should be correlated with clinical status of the patient
• Check the epidemiological situation

Question 10

ELISA (Enzyme-Linked Immunosorbent Assay)

Positive Result

Answer 10

Color Formation

Question 11

ELISA (Enzyme-Linked Immunosorbent Assay)

Types

Answer 11

• Solid Phase (test tube or microtiter plate)
• Membrane Bound (nitrocellulose membrane)

Question 12

Latex Agglutination

Positve Result

Answer 12

Agglutination

Question 13

Latex Agglutination

if the plasma contains anti bodies, what will happen

Answer 13

world stop

chz agglutination

Question 14

Electron Microscopy - TOF

10 to the 6th virus particle per mL is required for visualization

Answer 14

True

i also dk what shes talking abt <3 - this is abt the micrscope

Question 15

Electron Microscopy - TOF

electron micrscropy it can be made more sensitive, even at lower viral particles by using immunoelectron microscopy

Answer 15

T

Question 16

Electron Microscopy

Magnification:

Answer 16

50,000 to 60,000

Question 17

Cytology

uses what microscopy

Answer 17

Light Microscopy

Question 18

Cytology

what does this detect

Answer 18

changes in cells

Question 19

Cytology - TOF

checks for viral particles in cell

Answer 19

False

Question 20

Cytology

Rapid diagnosis of viral infections that produce??

Answer 20

Cytopathic Effects (CPE)

Question 21

Cytology

Characteristics of CPEs:

Answer 21

• Changes in cell morphology
• Cell lysis
• Vacuolation
• Syncytia
• Inclusion Bodies

Question 22

Cytology - TOF

Sensitive and Specific

Answer 22

False

Question 23

what are used under Classical Molecular Technique

Answer 23

• DNA/RNA probes for hybridization

Question 24

Classical Molecular Technique

what are under the DNA/RNA probes for hybridization

Answer 24

• DOT blot
• Southern Blot
• In-situ hybridization (DNA in cell)
• FISH

Question 25

# Classical Molecular Technique - TOF this techniques usually more tedious and expensive than the conventional techniques. Not widely accepted

Answer 25

True

Question 26

what are the newer molecular techniques

Answer 26

- PCR (amplification technique) - Commercial proprietary techniques

Question 27

# Newer Molecular Techniques under pcr

Answer 27

- Real-time PCR - RT-PCR

Question 28

# Newer Molecular Technique Commercial proprietary techniques

Answer 28

- NASBA: Nucleic Acid Based Amplification - LCR: Ligase Chain Reaction - TMA: Transcription-Mediated Amplification

Question 29

# Indirect Examination what is the first thing should be done

Answer 29

Virus Isolation

Question 30

# Indirect Examination after virus isolation, what step should incur

Answer 30

Virus Identification

Question 31

# Indirect Examination what are the can be used as a medium under viral isolation?

Answer 31

- Cell culture - embryonated eggs - chick embryo - seldomly used - animals can also be used

Question 32

# Indirect Examination what are the 3 types of Cell Cultures?

Answer 32

- Primary - Semicontinuos (Secondary) - Continous

Question 33

# What type of cell culture - From normal adult tissue (usually monkey kidney) - Can be subcultured (once or twice) - Best cell culture system (Supports widest range of viruses) - Very expensinve

Answer 33

Primary Cell Culture

Question 34

# What type of cell culture - From Embryonic tissue – Considered as stem cell (kaya pwede ma subculture ng marami) - Human embryonic kidney and skin fibroblast

Answer 34

Semicontinuous (secondary) cell culture

Question 35

# What type of cell culture - From Malignaant Tissue (HeLa Vero. Hep2 LLC-MK2 and MDCK) - Most easy to handle – cause its immortal cells - Easier to grow – grow in resit surface (??) and suspension - Can be subculture immortaly and indefinitely

Answer 35

Continous cell culture

Question 36

if u see this card

Answer 36

go over the cell culture

Question 37

what are the 2 factors to be considered in detection of the virus in culture?

Answer 37

- Presence of cytopathic effect - Hemeadsorption

Question 38

# Detection of the Virus in Culture - under what factor this maybe specific or non-specific - Balloning of cells - Syncytia formation - Detaching of cells in the plate

Answer 38

Presence of cytopathic effect

Question 39

# Detection of the Virus in Culture - under what factor Positive Result: Heme agglutination; carpet cell of red cell Negative Result: Button cells

Answer 39

Hemeadsorption

Question 40

what are the test under Conformation of the Identity of Virus

Answer 40

- Neutralization Test - Hemagglutination Inhibition (HAI) Test - Immunofluoresence

Question 41

# Conformation of the Identity of Virus (Virus + specific Ab against virus) + different cell culture → NO Cytophatic Effect

Answer 41

Neutralization Test

Question 42

# Conformation of the Identity of Virus (Virus + specific antibody) + RBC → NO hemagglutination

Answer 42

Hemagglutination Inhibition (HAI) Test

Question 43

Virus (place in a slide) + specific Fluorescein-labelled antibody → Fluoresence

Answer 43

Immunofluoresence

Question 44

A variation on standard tissue culture

Answer 44

Shell Vial Culture

Question 45

# Shell Vial Culture what is the technique

Answer 45

A small bottle with a removable round glass coverslip is used to grow the cells as a monolayer on the coverslip

Question 46

# Shell Vial Culture Result:

Answer 46

5-7 days | this is considered fast

Question 47

if u see this card

Answer 47

go over the transes and study the process

Question 48

also called as paired sera

Answer 48

Serology

Question 49

Detect antibodies but it takes two week or more before positive result are obtained

Answer 49

Serology

Question 50

# Serology - TOF also for detection of antigen in antibodies of viruses

Answer 50

True

Question 51

# Serology - TOF Detection of rising titers of antibody

Answer 51

True | being check is increase in titer of antibody

Question 52

# Serology Detection of ___ and ___ in primary infection

Answer 52

IgM (also IgG)

Question 53

# Serology checking of IgM ONLY

Answer 53

detect primary infection

Question 54

# Serology checking of IgM and IgG

Answer 54

positve for convalescent stage

Question 55

# Serology checking of IgG only

Answer 55

infection has been resolved

Question 56

what are the classic technique under Serology

Answer 56

- Complement Fixation - Hemaagglutination-Inhibition Technique - Immunofluorescent Technique - Neutralization - Counter Immunoelectrophoresis

Question 57

what are the newer technique under Serology

Answer 57

- Enzyme immunoassay - Particle Agglutination - Western Blot - RIBA (recombinant-immunoblot-assay) - Line immunoassay