SEROLOGICAL TESTING Flashcards
What is a non-treponemal test for serologic detection of the antibody Reagin?
Rapid Plasma Reagin
What is produced in response to an infection of syphilis caused by Treponema pallidum?
Reagin
What is a phospholipid, which is produced in patients infected with Treponema pallidum, the causative agent of syphilis, and acts like an antigen?
Reagin
Reagin is found in the serum/plasma of patients with what?
Syphilis
The vehicle or “antigen” used in an RPR is what?
Carbon charcoal particle
After the antigen is mixed with a serum/plasma sample in an RPR what would be a positive test?
If flocculation is present, the test is reactive (positive)
After the antigen is mixed with a serum/plasma sample in an RPR what would be a negative test?
If no flocculation is present, the test is non-reactive (negative)
What is the confirmatory test for RPR testing?
Fluorescent treponemal antibody-absorbed test (FTA-ABS)
What happens after the dead T. pallidum is fixed on a slide, and a patient’s serum added?
If antibody is present, organism is coated with antibody and is detected by addition of fluorescein-labeled anti-human globulin
What would a positive FTA-ABS test be?
This slide is studied under an ultraviolet microscope and will fluoresce if positive.
What would a negative FTA-ABS test be?
A negative test results shows no fluoresce.
What blood tube would you use for serum collection for an RPR?
Red/SST
What blood tube would you use for plasma collection for an RPR?
Purple top
True or False
After centrifugation, serum should be clear and non-hemolyzed
True
All of these supplies are included in what?
- RPR Card antigen suspension
- Plastic disposable bottle
- 20 gauge, galvanized needles, blunt tip
- Test Cards
- Pipettes/stirrers
- RPR Card Test Control Cards
- 1mL tuberculin syringe
- Distilled water
- Mechanical rotator
RPR Card Test Kit
True or False
Before testing for RPR you need to centrifuge the specimen (serum or plasma) to sediment cellular elements
True
How long should you mix the antigen suspension prior to opening the ampule?
vigorously shake for 10-15 seconds
What do you do after mixing the antigen suspension for the RPR test?
- Attach the needle to the hub of the dispensing bottle. Ensure the antigen is below the break line
- Snap off the top of the ampule and draw all of the antigen into the dispensing bottle
RPR Testing Procedure
Step 1
Collect specimen, hold a dispenser between thumb and forefinger. Squeeze, do not release pressure until open end is below surface of specimen. Holding tube vertically release finger pressure to draw up sample.
RPR Testing Procedure
Step 2
Use the dispenser to transfer the specimen to the card, dispense 1 drop.
RPR Testing Procedure
Step 3
Spread the specimen around the circle to cover the entire surface area within the circle.
RPR Testing Procedure
Step 4
Add one drop of antigen to each specimen and controls to be tested with the dispensing bottle.
RPR Testing Procedure
Step 5
Place test card and controls on the rotator for 8 minutes at 100 revolutions per minute (rpm).
RPR Testing Procedure
Step 6
Read the reaction at the end of 8 minutes.
RPR Testing Procedure
Step 7
Rotate and tilt card briefly, by hand (3 or 4 to-and fro-motion). This helps to differentiate non-reactive from weakly reactive results.
RPR Testing Procedure
Step 8
Examine test card. Read MACROSCOPICALLY, under a high intensity lamp or strong daylight.
RPR Testing Procedure
Step 9
Report results as either reactive or non-reactive.
RPR Reactive test results show flocculation ranging from what?
- *1. Slight but definite**
- *2. Minimum to moderate**
- *3. Marked and intense**
Reactive Rapid Plasma Reagin tests must be confirmed with what?
Fluorescent Treponemal Antibody Absorption test (FTA-ABS).
True or False
Rapid Plasma Reagin controls should always be run with each batch of Rapid Plasma Reagin tests.
True
What does quality control do ?
ensures the accuracy and reliability of test results
What are the two sections that quality control is divided into ?
- *1. Equipment quality control**
- *2. Reagent quality control**
Quality control equipment for RPR testing include what?
- *1. Mechanical Rotator**
- *a. calibrate to 100 rpm**
- *2. Antigen needle**
- *a. calibrate to 60 drops per 1 mL**
What are solutions that contain the same constituents as those being tested?
Control reagents
For Rapid Plasma Reagin what control reagents do you use?
- *1. Reactive**
- *2. Weak-reactive**
- *3. Non-reactive**
True or False
RPR control reagents are treated as patients and run every time an unknown sample is tested
True
What test is used to detect infectious mononucleosis which is a self limiting disease caused by the Epstein-Barr virus?
The Monospot
What disease is typically seen in young adults and children less than 5 years of age?
Infectious mononucleosis
What are these symptoms of?
- fever
- malaise
- lethargy
- sore throat with exudates
- enlarged lymph nodes on the neck
- mild hepatitis
- enlarged spleen
- occasional blotchy skin rash
Infectious mononucleosis
What causes the accumulation of increased numbers and abnormal forms of lymphocytes and monocytes in the lymph nodes, thus causing formation of “heterophile” antibody?
Infectious mononucleosis
What is an antibody that is structurally similar to the antibody that is specifically produced as a response to antigen stimulation
heterophile antibody
All of these supplies are included in what?
- Latex Reagent suspension
- Pipettes/stirrers
- Test slides
- Positive Control
- Negative Control
Monospot Latex Test Kit
Prior to Testing (Monospot)
Should you centrifuge the specimen (serum or plasma) to sediment cellular elements?
YES
What is the timeline to test monospot specimens if they have been stored between 2-8 degrees Celsius?
24 hours
When testing monospot should you use hemolyzed or contaminated samples?
….no….
True or False
You always perform quality controls, positive and negative, for monospot
True
Monospot Testing Procedure
Step 1
Allow the Latex Reagent and controls to reach room temperature (20 to 30 degrees C).
Monospot Testing Procedure
Step 2
Gently shake the Latex Reagent vial to disperse and suspend the latex particles in the buffer solution. Vigorous shaking should be avoided.
Monospot Testing Procedure
Step 3
Use the dispenser to transfer the specimen to the card, dispense 1 drop.
Monospot Testing Procedure
Step 4
Add one drop of Latex Reagent next to the drop of sample.
Monospot Testing Procedure
Step 5
Mix both drops with stirrer covering the whole surface of the slide section.
Monospot Testing Procedure
Step 6
Gently rotate the slide for 3 minutes manually or on a rotatory shaker set at 60-100 rpm.
Monospot Testing Procedure
Step 7
Look for the presence or absence of agglutination after the aforementioned period of time.
Monospot Testing Procedure
Step 8
Report results as either positive or negative.
Monospot Test Results
Positive result
Shows agglutination
Monospot Test Results
Negative result
Demonstrate a smooth, homogenous solution. No Agglutination.
What ensures the accuracy and reliability of test results?
Quality control
What starts with proper collection and handling of specimen, assaying control reagents, and checking instrument performance?
Quality Control
What are the two sections that quality control is divided into ?
- Equipment quality control
- Reagent Quality control
Quality control equipment for Monospot includes the what?
Mechanical Rotator
Calibrate to 100rpm