MEDT 25 | VIRTUAL INTERNSHIP AND SKILLS ENHANCEMENT

L4: Immunology and Serology Section

Lecturer: Anthony B. Dirain III, RMT
Transcribed by: MC, Centeno and KK, Colarina | 26 April 2023

Outline • track down the relevant T and B cells of the specific immune
I. Immunology system = activating specific immune system
II. Serology
III. Serology in EACMed Activate Complement System
• invading pathogens activate the complement system directly via
I. IMMUNOLOGY the lectin pathway and alternative pathway
• The Immune System is made up of:
o Organs Innate Immune System: Immediate Response Mechanism
o Tissue
• 1st response: macrophages
o Cells
o They recognize the pathogens by specific characteristics
o Molecules
that occur on pathogens, but don't occur on cells of the
• These generate an immune response to protect us from
body.
microorganisms, remove toxins and destroy tumor cell.
o These characteristics are called pathogen-associated
• Immune response can (all within 10 days): molecular patterns or PAMPs
o Identify a threat o They recognize PAMPs using various receptor types,
o Mount an attack including toll-like receptors.
o Eliminate pathogen
o Develop mechanism
PHAGOCYTOSIS
• Prolonged reaction = chronic inflammation
• process that macrophages and other phagocytes like neutrophils
use in order to destroy pathogens
Types of Immune System
Phagocytosis: Process
1. Once they recognize the pathogen, macrophages start to wrap
their cell membrane around the pathogen and absorb them with
inside their cells.
2. Pathogen is then left inside the phagosome.
3. This phagosome fuses with surrounding lysosomes, which provide
digestive enzymes that then break down that pathogen,
destroying it and also processing all the components into
harmless waste products.

BARRIERS
Physical Barriers
• SKIN
o There are physical barriers that need to be broken before
an infection can take place.
o The skin is an example of the barrier, just invaders will be
stuck at this barrier.
• MUCOSA (respiratory, gastrointestinal, urinary)
INFLAMMATION
o However, an invading pathogen will get through, such as
when there is a cut in the skin • The invading pathogens are harmless enough that macrophages
can deal with them alone. They are cleared and it goes no further.
Chemical Barriers • If the attacking army (pathogens) is too many, macrophages may
need help.
• Chemical barriers that help destroys harmful pathogens before • Macrophages release cytokines
they can cause any infection such as the: o Cytokines – are signaling proteins like local hormones that
o Hydrochloric acid in the stomach sounds the alarm of an infection in the local area leading
o Lysozyme in sweat to a process called inflammation.
o Tears
o lactic acid in the vagina

INITIAL IMMUNE RESPONSE

• Once an invading pathogen has broken through the physical and
chemical barriers to infection, things will happen.

Macrophages
• recognize the pathogen
• activate the innate immune system
CYTOKINES
Dendritic Cells • Most important actions of these cytokines is to recruit and
• pick up antigens from the pathogen and then head off in the activate more cells of the immune system:
blood and lymphatic systems
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 Lesson 4: Immunology and Serology

o Macrophages – differentiated monocytes when it entered • marker of inflammation

the tissue • can be a really good indication of the severity of the infection
o Monocytes – are precursors to macrophages that float that the patient is suffering with
around in the blood
o Neutrophils – are another type of phagocytes that SUMMARY OF INNATE IMMUNE SYSTEM
circulate in the blood and can enter the tissues and help • Generalized system that recognizes and responds an invading
by destroying invading pathogens pathogen through:
o Inflammatory response
INFLAMMATORY RESPONSE o Recruiting cells that destroys pathogen by phagocytosis
• A number of processes that help to contain and fight infection:
o vasodilation
o vascular permeability
o mast cell degranulation – releasing more cytokines that
further stimulates the inflammatory response
o activation of clotting and kinin system

Acute Phase Response

• Inflammation itself stimulates macrophages and neutrophils to
secrete more cytokines notably called interleukins.
o Interleukins – known as the acute phase response and it
leads to a more substantial response
COMPLEMENT SYSTEM
• The complement system works alongside the innate and specific
immune system to help them destroy pathogens.
• It involves a series of complement proteins labeled C1 to C9.
• Once it is triggered, proteins start to activate each other in
something called complement cascade.
• Various products (C3a, C3b, C4a, C5a, MAC) of the complement
cascade have important functions such as:
o acting as opsonin
o triggering further inflammation
o directly attacking and destroying pathogens.

Interleukin-1
• Inflammatory response involves sending cytokines to the brain to
tell it to produce a fever that leads to high temperature that are
poorly tolerated by many pathogens
• this causes reduce appetite and lethargy so that the person
conserves more energy that can be used to fight infection

Interleukin-6
• Inflammatory response involves sending interleukin six to the
liver to produce acute phase proteins that act as something
called opsonin.
Pathways of Complement System
Interleukin-8
• its release recruits and activates more neutrophils • Complement system is triggered in one or three ways.
o Lectin Pathway
activated directly by pathogens
Interleukins-1 and 12 o Alternative Pathway
o Classical Pathway – activated by antigen antibody
• activates natural killer cells
complexes that arise from this specific immune system
TNF-alpha
• When tumor necrosis factor (TNF-alpha) is released it does all
things IL-1, IL-6, IL-8, and IL-12 are doing.

OPSONINS
• complex molecules that attach themselves to pathogens
• it is easier for macrophages and neutrophils to recognize and
phagocytose the pathogen because of opsonin
• Example: C-reactive protein (CRP)

C-Reactive Protein (CRP)

• produced by the liver in response to interleukin six
• measures the level of CRP and patients to assess how much
inflammation there is in the body

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 Lesson 4: Immunology and Serology
SPECIFIC IMMUNE SYSTEM
• Involves two characters: T and B cells
• These are both types of lymphocytes.
• They are free to float around the lymphatic system in the blood.
• They spend most of their time in the lymph nodes and in the
mucosa-associated lymphoid tissue.
Lymph Nodes
• Lymph nodes are like army barracks full of soldiers that are all
set around waiting to get a word for an enemy that they
specifically been trained to fight
T cells and B cells
• Each pathogen has molecules that are unique to them known as
antigens.
• Each T cell has T cell receptors that are specific to a single type
of antigen.
• B cells have antibody on their cell membrane, just like T cell
receptors that are specific to a single type of antigen.
• Result: there are millions of different T and B cells that are all
specific to a single type of antigen
Specific Immune System: Response Mechanism
• When a new pathogen arrives and causes an infection, the T and
B cells that are specific to that pathogen need to be alerted.
• The trouble is there are certain lymphatic tissues such as lymph
nodes, while the infection is happening somewhere completely
different.
• This is where the dendritic cells come in.
o Dendritic cells – are like messengers and they pick up the
antigens at the site of invasion and display them through
the blood of the lymphatic system into the lymphatic tissue.
• When dendritic cells are there, all the T and B cells have a look at
the antigen and see whether they recognize it.
• When the T and B cells are found that this specific to the antigen
those cells become activated.
T-CELL DIFFERENTIATION
• T-cells can differentiate into: T-helper cells
o T-helper cells can differentiate into:
▪ Cytotoxic T-cells
▪ Plasma Cells and Memory B cells
▪ Macrophage activator
T-cell Differentiation: T-helper Cells
• Specific immune response starts in the dendritic cells presenting
the antigens on their HLA Class two molecules to the CD4 cells.
• These CD4 cells, which are a type of T cells then proliferate and
become T helper cells.
• T helper cells present antigens on their HLA class, one molecule
that can be recognized by CD4 cells, and another type of T cell.
T-cell Differentiation: Cytotoxic T-Cells
• T-cells also secrete cytokines where it's possible or make CD4 cells
proliferate and differentiate into cytotoxic T cells.
T-cell Differentiation: Plasma cells and Memory B cells
• T helper cells also release cytokines that stimulate B cells to
proliferate and differentiate into plasma cells.
• Plasma cells can release large quantities of antibodies and
memory B cells that hang around as part of the immune memory
to respond quickly in future infections with that specific
pathogen.
T-cell Differentiation: Macrophage Activator
• The T helper cells also traveled to areas of infections and secrete
cytokines that help to recruit monocytes and macrophages to the
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 Lesson 4: Immunology and Serology
infected tissue and activate the macrophages to cause
inflammation and start the process of phagocytosis.
T-cell Differentiation
CYTOTOXIC T-CELLS
• Cytotoxic T cells are responsible for killing cells that have been
infected by pathogens such as virally infected cells.
• To do this, they need to attach themselves to the infected cells
via T-cell receptor and HLA Class I molecule expressing the
relevant antigen protein on the infected cell.
2 Killing Mechanisms of Cytotoxic T-cells
1. Granule exocytosis – where they basically spray the infected
cell with enzymes that destroys the membrane and lead to cell
lysis and cell death
cell death
2. Activation of FAS molecule – is like a “self-destruct switch” that
once it's activated, it causes the cell to undergo apoptosis.
apoptosis
PLASMA CELLS AND ANTIBODIES
• PLASMA CELLS AND ANTIBODIES are an essential part of the
specific immune system
• Plasma cells or B cells – have differentiated and become
antibody producing cells and their job is to produce antibodies
that are specific to the invading pathogen.
Antibodies
• proteins that are shaped like “Y”
• one end is variable in shape to match different antigens
• whereas the other end is fixed in shape and can recognize by
many cells of the immune system
• Antibodies float around the blood and attach themselves to
antigens that match their specific variable region.
Antibody Structure
Antibodies helps the immune by pathogens in a number of
different ways:
1. They can attach themselves to enemy toxins which themselves are
antigens and neutralizes their toxic effect.
2. Antibodies can attach themselves to the receptors of viruses and
bacteria, and prevent them for carrying out their function. For
example, it can stop viruses from being able to recognize cells
that it may want to invade, and therefore prevent the viral
invasion of the cell.
3. Antibodies can attach themselves to the pathogens they clump
together to slow the spread of that pathogen down. This is called
agglutination.
4. It can be very difficult for the simplistic receptors of
macrophages and neutrophils to recognize certain pathogens.
Antibodies can act as opsonin that are highly specific to the
invading pathogen. By attaching themselves to that pathogen
acting as an opsonin, they can help the macrophages and
neutrophils to recognize and destroy that pathogen.
II. SEROLOGY
• Scientific study of serum and other body fluid in practice
• term usually refers to serologic test
Serologic Test
• Blood tests that look for antibodies in your blood that can involve
a number of laboratory techniques.
• Different types of serologic tests are used to diagnose various
disease conditions.
• Serologic Tests have one thing in common, they all focus on
proteins made by your immune system. This vital body system
helps keep you healthy by destroying foreign invaders that can
make you ill.
• The process of having the test is the same regarding which
technique the laboratory uses during serologic testing.
Why do we need a serologic test?
• It’s helpful to know a little about the immune system and why we
get sick to understand serologic tests and why they are useful.
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 Lesson 4: Immunology and Serology

ANTIGENS MACHINE AND KITS

• Antigens are substances that provoke a response from the 1. Maglumi 800
immune system. 2. Finecare FIA meter
• They're usually too small to see with the naked eye.
• They can enter the human body through the mouth, through A. MAGLUMI 800
broken skin, or through the nasal passages. • New member of Snibe Maglumi series
• Antigens that commonly affect people include the following: • Fully automated
o bacteria • Chemiluminescence immunoassay analyzers (CLIA)
o fungi 1. Labeling technology - determines reaction mode
o viruses 2. Separation technology - determines sensitivity, accuracy,
o parasites and precision of reagents

How does the immune system fight antigens?

• The immune system defends against antigens by producing
antibodies.
• These antibodies are particles that attach to the antigens and
deactivate them.
• When your doctor tests your blood, they can identify the type of
antibodies and antigens that are in your blood sample, and Maglumi 800
identify the type of infection you have.
Advantages:
AUTOIMMUNE DISORDERS • For small and middle size lab or hospital
• Sometimes the body mistake its own healthy tissue for outside • Simple, smart, humanization-designed model
invaders and produce unnecessary antibodies • More than enough test menu to meet the different needs and it
• This is known as an Autoimmune disorder. is enlarging according to your requirement
• Serologic testing can detect these antibodies and help doctor to • Free QC and calibrator help to reduce the cost per test
diagnose an autoimmune disorder. • Key technology ABEI and Nanomagnetic microbead enhances
stability and sensitivity of MAGLUMI reagents
TYPES OF SEROLOGIC TEST
• Antibodies are diverse, so there are various tests for detecting Parts of Maglumi 800
the presence of different types of antibodies. 1. Pipettor - aspirate reagent and sample with high speed
• These include: o Titanium needle for crush proof
1. Agglutination Assay – shows whether antibodies exposed o Clot detection and liquid level detection
to certain antigens will cause particle clumping o Coated with Teflon to prevent carry over
2. Precipitation Test – shows whether the antigens are similar o Inner and external washing
by measuring for the presence of antibody in body fluids 2. Automate cuvette loader- up to 40 modules of 240 tests and
Continuous loading
NORMAL VS ABNORMAL TEST RESULTS 3. Sample area- up to 40 samples on board with barcode and
NORMAL RESULTS ABNORMAL RESULTS Continuous loading and cooling function
• Body produces antibodies • Antibodies in blood sample 4. Incubator- incubate 13 slots (78 tests at the same time), temp.
in response to antigens. often mean you have had 36.8C
• If testing shows no an immune system response 5. Reagent area- up to 9 reagents on board with RFID and easy
antibodies, it indicates you to an antigen from either and fast for adding and updating
do not have an infection. current or past exposure to 6. Washer- strong magnetic field helps betted separation and 3
• Results that show there are a disease or foreign protein. pairs of washing needles with antioverflow function
no antibodies in the blood • The presence of certain 7. Chamber- high sensitivity and low-noise PMT and cuvettes
sample are normal. types of antibodies can also detection and anti-overthrow
mean that you are immune
to one or more antigen. This
means that future exposure
to the antigen/s won’t
result in illness.

What happens after serologic testing?

• The care and treatment provided after serologic testing can vary.
• It often depends on whether antibodies were found.
• It may also depend on the nature of your immune response and
its severity.
✓ An antibiotic or another type of medication may help your
body fight the infection. Even if your results were normal,
the doctor might order an additional test if they still think
you might have an infection.
✓ The bacteria, virus, parasite, or fungus in your body will Tests Available (Thyroid function tests)
multiply over time. In response, your immune system will T3 (triiodothyronine) & T4 (thyroxin)
produce more antibodies. This makes the antibodies easier • Helps to control how your body stores and uses energy or
to detect as the infection gets worse. The test results may metabolism
also show the presence of antibodies related to chronic • Help control many of our body's other processes.
conditions such as autoimmune disorders. o Breathing

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 Lesson 4: Immunology and Serology

o Heart function 3. Next to the reagent area, there is a sensor that automatically
o Nervous system function reads chips on reagent packages and passes on the information
o Body temperature to the software
o Cholesterol level how easily you gain weight 4. With SNIBE technology, there are more than 120 kinds of reagent
o Brain development for choosing
o Moisture of the skin 5. Maglumi reagent kinds contain a magnetic microbead suspension
o Menstruation. rotor
6. These beads are suspended in solution via the automatic mixing
Normal Value that occurs after loading
T3 0.69- 2.15 ng/ml 7. In regards to the maglumi 800, it has a capacity for 9 reagents
T4 52-127 ng/ml and can be continuously loaded
8. Samples can be loaded with different types of sample tube
9. When loading the sample, the barcode reader scans the barcode
This test can show your healthcare provider if your direct gland is on the tube and identifies the sample ID sending such information
overactive or underactive. onto the software
10. The maglumi 800’s sample area has a maximum capacity of 40
TSH, FT3 & FT4 samples and can continuously load samples
• Aids in the diagnosis of thyroid disorders. 11. The reagent and sample are also refrigerated which helps increase
o TSH - serves as an initial test and thyroid diagnostics. stability
o FT3 - provides a further confirmatory test for monitoring 12. Maglumi software can automatically obtain the worklist via
laboratory information system
hyperthyroidism to supplement the T3 and T4 in thyrotropin
13. A multi-language and user-friendly interfere also facilitates the
assays
operation
o FT4 - measured in response to abnormal TSH Test Results
14. After confirmation of the worklist, click “Start” button to start the
test
Normal Value 15. As the test commences, the sample needle pipettes the sample and
TSH 0.3-4.5 uIU/ mL reagent in the cuvette
FT3 2.0- 4.2 pg/ mL 16. The sampling needle is titanium a TEFLON coated which makes it
FT4 8.9- 17.2 pg/ mL crush proof
17. Internal and external washing plus liquid and clot detection all
INFLAMMATORY MONITORING ensure accurate pipetting
18. After the pipetting is complete, the cuvette is transferred to the
Procalcitonin (PCT)
incubator for 10 to 15 minutes
• Procalcitonin is a biomarker associated with the inflammatory 19. The incubator has 13 incubation slots available and has precise
response from bacterial infection that aid in the risk assessment temperature control to 37C
of critically ill patients on their first ICU admission 20. Following incubation, the cuvette is moved into the washing
o Patients on their first day of ICU admission for progression station
and septic shock. 21. Here the magnetic micro beads are attached to the magnetic
o It also helps guide an antibiotic treatment and or monitor positioned at the back of the cuvette
the effectiveness of treatment. 22. Cuvette is then cleaned 3 times with wash buffer to remove
unconjugated contents
Normal Value 23. One bottle of concentrated wash buffer is generally diluted to
Procalcitonin (PCT) <0.5ng/L 10uL for this purpose
24. Finally, the cuvette moves into the measuring chamber which is a
darkroom environment
Tumor Markers 25. Here, 2 substrates: Starter 1 and Starter 2 are added. They react
with present chemiluminescent label ABEI
Tumor Marker Uses Normal Values 26. After measuring, the cuvette is pushed into the waste bag and the
Used in the evaluation of test ends
Ferritin iron deficiency and iron 13-350 ng/mL
overload Operation Overview
Aids in the detection of 1. The user loads the desired 2. The samples are loaded
Prostate-
prostate cancer in men < 4.0ng/mL reagents
specific
antigen (PSA) aged 50 years or older

Infectious Disease
Anti-HBs
• Used to monitor the success of Hepatitis B vaccination
• Used to monitor convalescence and recovery of Hepatitis B
infected individuals 3. Desired Worklist is input 4. Press “Start” and wait for
• Cutt-off value: 10mIU/mL manually the results

Note: Running is performed by Medical Technologist with

proficiency training

Operation
1. Maglumi 800 can load up to 40 pcs of cuvette at a time which can
run 240 tests. During testing, these can continuously be added
2. Maglumi kit uses RFID technology

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 Lesson 4: Immunology and Serology

B. FINECARE FIA METER

• Fluorescence immunochromatographic analyzing system which
help diagnose condition such as infection, diabetes,
cardiovascular diseases, renal injury and cancers
• The finecare platform provides reliable and quantitative results
of various kinds of analytes in human blood or urine within
several minutes 9. Insert the cartridge in the 10. Press “IN” button and
test channel “Test” display to start
Parts of Finecare FIA Meter the test
1. Built-in thermal printer – for real time printing
2. LIS/ HIS connection – direct connection to LIS/ HIS
3. ID Chip port – contains all information of test item and lot
number
4. Test Channel – Test cartridge holder
5. LCD – 3.5 LCD touch screen
11. Wait 3 mins for the result 12. Input corresponding
accession number and
press “OK” button

13. Click “Print” button to print. Record your results

Test Available
C-reactive protein (CRP)
• a C-reactive protein test measures the level of c-reactive protein
in your blood.
o CRP is a protein made by your liver. It’s sent into your
bloodstream in response to inflammation
o Inflammation is your body’s way of protecting your tissues
if you’ve been injured or have an infection – It can cause Note: The ID chip contains test item, batch number, standard
pain, redness, or swelling in the injured/ infected area. graph; after finishing test. All information in ID chip was stored in
▪ Some autoimmune disorders/ chronic diseases can also the equipment, thus it is not necessary to insert ID chip of the
cause inflammation same batch again
o Normally, low levels of C-reactive protein are found in
blood. High levels maybe a sign of serious infection or other TEST KITS
disorders 1. SDBIOLINE DENGUE DUO RAPID TEST
o Cut-off value: <10ng/mL
• Designed to detect both dengue virus NS1 Antigen and
differential IgG IgM antibodies to dengue virus
Operations
• SD Dengue NS1 Antigen could be useful for early acute phase
1. Prepare all materials 2. Insert ID Chip
samples from day one to five
needed
• An IgG and IgM antibody test was designed to detect
antibodies appearing in the convalescence phase after day 5 to
14
• Main purpose of the test is the simultaneous detection of dengue
NS1 antigen and IgG and IgM antibody together, not separately.

Advantages
3. Remove test device from 4. Pipette 5 ul of serum or • Diagnosis of Dengue NS1 antigen even during window period
the foil pouch 8.5 uL of whole blood • Compensate for the weak point of Antigen single test
• Fast test result: (15-20 minutes)
• Convenient storage condition (1-30C)

Materials Included
• Test pouch
5. Mix with CRP buffer 6. Invert for 1 minute
• Buffer
• Disposable dropper
• Capillary pipette
• Package insert

Note:
7. Transfer 75uL of mix 8. Click “Standard Test” and • All test materials must be used immediately once opened
buffered and serum into press “Out” display procedure
the test kits button

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 Lesson 4: Immunology and Serology

• DO NOT read the test result after 20 mins reading too late 2. SD BIOLINE HIV 1 AND 2 RAPID TEST
can give FALSE result • Designed for differential detection of all isotype antibodies
specific to HIV type 1 including subtype O and type 2
Procedure simultaneously
1. Check the expiry date on the backside of the test pouch
o Note: If the expiry date has passed, use another kit Advantages
2. Remove the test device from the foil pouch and place it on the • Serum, plasma or whole blood can be used as specimen
flat surface • Test result is only 20 mins
3. Write the corresponding accession number in the test kit • Storage condition is RT for 2 yrs
o Note: Always check patient’s request and sample if its • Sensitivity is 100% and specificity 99.8%
correct
4. For NS1 testing, using a disposable dropper provided, take serum
Materials include
or plasma
5. And Add 3 drops of serum or plasma into the sample well marked • Test pouch
as “S”’ • Buffer
6. At the same time, using the capillary pipette, take sample from • Package insert
the collection tube • Capillary pipettes
7. And drop 10 microliters of serum or plasma into the sample well
for antibody testing Note: All test materials must be used immediately once opened
8. Put 4 drips of diluent into the round-shaped assay well
9. As the test begins to work, you will see purple color move across
Procedure
the result window in the center of the test device
1. Check the expiry date on the backside of the test pouch
10. Interpret test results after 15-20mins.
o Note: If the expiry date has passed, use another kit
2. Open the test ouch and set the test device
Note: DO NOT read the results after 20 mins. Reading too late can 3. Write the corresponding accession number in the test kit
give FALSE results 4. Using a micropipette take 10 microliters of serum or plasma
5. And put it into the round sample well
Interpretation of Results 6. Add 4 drops of buffer into the same round well
Negative NS1 Antigen Positive 7. Read the results in 20 mins.

Note: DO NOT read the results after 20 mins. Reading too late can
give FALSE results

Interpretations
One line “c” on result window Indicative of acute stage Negative HIV 1 Positive
dengue infection
NS1 Antigen/ IgM Positive IgM Positive

One line “c” on result window

Two color bands (hiv-1 test line

Indicative of acute stage and “c” control line) or three color
Primary dengue infection
dengue infection bands (hiv-1 test line, “c” control
line, and faint “hiv2” test line)
IgG Positive IgG/ IgM Positive HIV 2 Positive HIV 1 and 2 Co-Infection

Perform confirmatory test

Secondary/ past dengue

infection Two color bands (“HIV-2” test line
Invalid Result and “C” control line) or three color
bands (HIV-2 test line, “c” control
line, and faint “HIV-1” test line)
Invalid Result

No “c” line repeat test using a new test device

Each test can be used ONCE do not try using the test more than No “c” line on result window, repeat using a new test kit
once.

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 Lesson 4: Immunology and Serology

3. SD BIOLINE SALMONELLA TYPI IG AND IGM FAST INTERPRETATION OF RESULTS

• Is an immunochromatographic assay for the rapid, qualitative, Reactive Invalid
and differential test for the detection of IgG and IgM antibodies
to Salmonella Typhi in human serum, plasma of whole blood
• This test provides only a preliminary test result. Therefore, a more
specific alternative diagnosis method must be used in order to
confirm Salmonella typhi infection

Benefits
• IgM: perfect alternative of Widal test presence of bands on control no band formation or band on
and patient lines the test line but not band on
• IgG: an examination of past infection
• Test result: 15-30 mins control line
• Storage condition: 2-30 C for 24 hr.
RAPID PLASMA REAGIN (RPR)
Note: DO NOT read the test result after 30 mins reading too late • Before, RPR was used in EACMed for detection of syphilis, but now
can give FALSE result immunochromatography assay is used.
• Principle: Macroscopic flocculation
• Test type: Screening test for syphilis but not specific (rapid test)
Materials
• Target analyte: Reagin
• Test Pouch
• Carrier: Charcoal particles
• Diluent
• Specimen: plasma and serum
• Package insert
• Disposable test tube
INTERPRETATION OF RESULTS
• Loop
Reactive Nonreactive
Procedure
1. Check the expiry date on the backside of the test pouch
o Note: If the expiry date has passed, use another kit
2. Open the test pouch and set the test device
o Note: Allow the kit to come to room temperature before
testing
3. Dispense 4 drops of assay diluent to the disposable test tube
4. With the loop provided, dispense 1 µL of the specimen to the test presence of flocculation no flocculation
tube containing assay diluent
o Note: Dip the circular end of the loop into the specimen, and ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)
then carefully place the circular end of the loop into the test • method for Hepa B and HIV test
tube. This will add 1ul of specimen to assay buffer • first ELISA is radioimmunoassay
5. Gently stir the assay buffer with the loop to ensure adequate • used to detect either antigen or antibody
mixing of specimen in the assay buffer • conjugate (enzyme) attaches to target analyte – Ag or Ab
6. Holding the strip vertically, insert the test strip into the tube
• substrate catalyzes the enzyme to form a color change or
containing diluted specimen
fluorescence
7. Interpret test results at 15-30 minutes
• Optical density (OD) is then measured by a machine; cut off
o Note: If test band is very faint at 15 minutes, read the
calculation follows
results again at 30 mins.

Note: DO NOT read the test result after 30 mins reading too late
can give FALSE result

SEROLOGY IN EACMED
• Two methods are used for testing STIs in EACMed:
1. Immunochromatography Test
2. Enzyme-linked Immunosorbent Assay (ELISA)

IMMUNOCHROMATOGRAPHY TESTS
• Immunochromatography assay
ELISA Strip and Strip holder
• Lateral flow assay
• Purpose: used to detect target analyte Machines
• Specimen: plasma, serum, whole blood
• EIA Reader = URIT-660
• rapid diagnosis (average of 15 minutes)
• Screening Test: HIV, HBsAg, Anti-HCV, Syphilis

URIT-660

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 Lesson 4: Immunology and Serology

• Washer EXTERNAL QUALITY ASSURANCE PROGRAM / SCHEME

• reference laboratories are sending samples for testing
• results are encoded in the websites of the reference laboratories
within the due date
• certificates are given to laboratory for participating in EQAS
o Certificate of Participation – failed EQAS
o Certificate of Proficiency – passed EQAS

Why is EQA important?

• evaluate the overall performance of the laboratories
• Incubator – should always be at 37°C • identify problems
**do not proceed to testing if temperature not achieved** • collect information
• establish information exchange network

EQA should lead to corrective action

• Corrective action – action taken to correct a problem or
deficiency

IDENTIFY TAKE CORRECTIVE

EQA
PROBLEMS ACTION

ADDITIONAL NOTES:
INTERPRETATION OF RESULTS
• When a blood unit is reactive for screening test, it is placed
Reactive OD reading above the cut-off value. under quarantine. The papers are prepared to be passed on
Nonreactive OD reading below the cut-off value. the reference laboratories together with the blood unit. The
blood unit is transported to RITM, then result will be released
Note: Cut-off value is dependent on the optical density of the after 2 weeks.
negative control. • According to DOH, the patient should be informed of the
reactive test results for TTIs. Then, the results is endorsed to
At Risk Populations Present in the Philippines (HIV) IDS (Infectious Disease Specialist).
• RITM receives positive blood units for TTIs.
• female sex workers
• STD/AIDS Cooperative Central Laboratory – San Lazaro
• male sex workers
Hospital (SACCL-SLH) tests individuals positive for STIs
• males who have sex with other males screening test.
• injecting drug users • STIs and TTIs have separate EQAS.
• clients of sex workers o STI EQAS samples will be given by SACCL-SLH
o TTI EQAS samples will be given by RITM
What puts Filipinos at risk for HIV?
• unprotected sex with > 1 partner
• injecting drugs with used needles
• mother was HIV+
• transfusion

Acquiring HIV from transfusion has a very low chance because

blood units are tested for Transfusion Transmissible Infections
(TTIs) before it is released.

Transfusion Transmissible Infections (TTIs)

• HIV
• Hepatitis B
• Hepatitis C
• Syphilis
• Malaria

QUALITY ASSESSMENT PROGRAMS (PROFICIENCY TESTING)

• Verify that laboratories are proficient in their testing process
(accurate results).

2 Types of Quality Assurance Programs

1. IQAP / IQAS – Internal Quality Assurance Program / Scheme
2. EQAP / EQAS – External Quality Assurance Program / Scheme

INTERNAL QUALITY ASSURANCE PROGRAM / SCHEME

• the quality assessment is conducted within the laboratory

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