1. Booth, Wallace, and Fitzgerald
Chapter 5
PowerPoint ® Presentation to Accompany
Phlebotomy for Health Care Personnel
Booth, Wallace, and Fitzgerald
Chapter 5

2. Specimen Handling and Processing
Specimen Processing
Specimen Handling and Processing
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3. Objectives Learning Outcomes
Upon completion of this chapter, you should be able to:
List requirements for special specimen handling procedures.
Describe the technique required for collecting blood cultures.
List the steps for Unopette® collection.
Describe the procedure for performing a microhematocrit.
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4. Learning Outcomes (cont'd)
Objectives cont’d
Learning Outcomes (cont'd)
Explain the collection process and procedure for making a peripheral blood smear.
Discuss common point-of-care and CLIA waived tests.
Explain the procedure for urine specimen collection and reagent testing.
Identify safety requirements for operating a centrifuge.
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5. Special Handling Procedures
Special Specimen Handling Procedures
In addition to blood collection, some specimens require special handling either before or after the blood is collected.
Common Tests Requiring Special Handling Procedures
Blood
Cultures
Legal Specimens
Cold Agglutinins
Chilled and Light-Sensitive
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6. Blood Cultures Blood Cultures Requested on patients with a high fever
Purpose is to isolate microorganisms from the patient’s blood that may be causing the fever
Strict aseptic technique is required for the collection of the blood specimen
Blood culture bottles are larger than normal venipuncture tubes
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7. Blood Cultures Part 2 Blood Cultures
Blood culture bottles contain culture media, which enhances the growth of the microorganisms
Septicemia is the presence of microorganisms in the blood with symptoms that can result in death
Blood cultures are usually ordered STAT
Collected in sets of two or three
Each set is collected from different sites at different time intervals
One sample will be incubated as an anaerobic specimen and the other as an aerobic specimen
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8. Blood Cultures Part 3 Blood Cultures Bacteria Fungi Protozoa Viruses
Blood cultures seek to identify the agent(s) responsible for the patient’s illness
Common Pathogen Classifications
Bacteria
Fungi
Protozoa
Viruses
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9. Blood Culture Volume Blood Cultures
The exact amount indicated by the manufacturer must be collected.
Usually 8 to 10 mL per bottle or tube is sufficient for an adult. Lesser amounts are drawn on infants and children.
The correct amount is needed to increase the chance of bacterial growth.
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10. Blood Cultures Part 4 Blood Cultures
The collection of blood cultures is essentially the same as for routine venipuncture, except:
Special site cleansing
Amount of blood needed
Specimen transfer
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11. Site Cleaning Procedure
Blood Cultures
Blood Culture Site Cleaning Procedure
Correct cleaning of the site is the most important part of collecting blood cultures
Steps to Correct Cleaning
Release the tourniquet after site is selected
2. Clean first with 70% to 95% alcohol, chlorhexidine gluconate, or benzalkonium chloride using concentric circles, from the center working outward
3. Do not allow the strokes to go back toward the center area
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12. Site Cleaning Procedure Cont’d
Blood Cultures
Steps to Correct Cleaning (cont’d)
4. Allow the site to air dry
5. Repeat cleansing using a 2% iodine swab or applicator if using alcohol
6. Allow iodine to dry and do not retouch the area
If the patient is allergic to iodine, use another cleanser.
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13. Blood Culture Specimen Transfer
Collecting Blood Culture Specimens
If using a syringe, be sure to have a syringe that can hold at least 20 mL. Attach the syringe to a transfer device and fill anaerobic bottles first.
If a butterfly setup is used, the aerobic sample is drawn first so that air from the butterfly tubing is cleared before the anaerobic culture sample is drawn.
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14. Legal Specimens
Establishing a chain of custody is required for medicolegal issues. Without proper proof of custody, the specimen will be considered invalid.
A chain of custody form must be completed correctly, and multiple copies are used as a safeguard system.
These specimens must be correctly identified and under the uninterrupted control of authorized personnel to ensure their validity.
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15. Cold Agglutinins Cold Agglutinins
Testing for cold agglutinins or antibodies is done for patients suspected of having atypical pneumonia.
Atypical pneumonia is caused by the microorganism Mycoplasma pneumoniae.
People infected with Mycoplasma pneumoniae produce autoantibodies.
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16. Cold Agglutinins Part 2 Cold Agglutinins
Normal body temperatures range from 97.6F to 99.6F. At temperatures lower than normal body temperature, cold agglutinins attach to red blood cells and cause clumping.
Specimen Collection
Use red-topped tubes that do not contain additives
Collection tubes must be prewarmed
Keep the sample at 98.6F until the serum can be separated from the cells
Serum must be separated from cells within 1 hour
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17. Chilled and Light-Sensitive Specimens
Some specimens must be chilled or covered immediately after collecting.
Chilled Specimens
Arterial blood gases
Ammonia
Lactic acid
Light-Sensitive Specimens
Bilirubin
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18. Apply Your Knowledge Part 2
Which of the following best describes the correct cleansing procedure for blood culture collection?
A. Clean the arm starting a distance from the site and clean toward the selected site
B. Use linear strokes to clean up and down the arm until the entire arm is cleansed
C. Clean the site starting in the center and working outward
Answer: C. Clean the site starting in the center and working outward
GREAT!
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19. Apply Your Knowledge Part 3
James is suspected of having atypical pneumonia. Which of the following tests will most likely be ordered?
A. Arterial blood gases
B. Cold agglutinin
C. Bilirubin
BRAVO!
Answer: B. Cold agglutinin tests for atypical pneumonia caused by Mycoplasma pneumoniae
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20. Unopette Procedure Unopette Procedure
A Unopette®, manufactured by Becton-Dickinson, is a prepackaged microcollection device.
Disposable
Consists of a plastic reservoir containing a premeasured volume of reagent
Common Tests Performed with a Unopette
White blood cell count
Red blood cell count
RBC fragility
Platelet count
Hemoglobin
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21. Unopette Procedure Part 2
Each test requires a specific Unopette that has a different premeasured volume of reagent inside.
Specific pipettes accompany each Unopette.
Pipettes are self-filling, thin-walled, glass capillary tubes attached to a plastic holder.
Use a dermal puncture specimen to fill the pipette.
Wipe the pipette after it fills with blood, but do not touch the end of the pipette with gauze.
Gently invert to mix the sample with the reagent.
Label Unopette and transport to lab.
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22. Apply Your Knowledge
Which tests can be performed with the Unopette microcollection device?
ANSWER: WBC count, RBC count, RBC fragility, platelet count and hemoglobin
Good Job!
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23. Measuring Microhematocrit
Microhematocrit tubes are used for measuring hematocrit or packed cell volume
They have a thin red band around the end of the tube
These tubes are coated with sodium heparin
Blood flows freely into these tubes because of capillary action
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24. Measuring Microhematocrit Part 2
Fill two capillary tubes 3/4 full
Seal the end that was not used to collect the specimen by embedding the clean end into a clay sealant (if the tube requires it)
Place in a separate tube for labeling and transporting to the laboratory
Place in centrifuge machine according to procedure
NOTE: The values of the two microhematocrits should match within 2 percent.
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25. Apply Your Knowledge Part 6
True or False: Microhematocrit tubes are coated with an anticoagulant.
Hey, remember! An
anticoagulant prevents
blood from clotting.
Great!
Answer: True
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26. Peripheral Blood Smear
A blood smear is for the microscopic examination of blood.
Venous or capillary blood may be used to prepare a blood smear.
The complete blood count (CBC) differential test is done with a blood smear.
Blood smears are also used to diagnose malaria, anemia, and leukemia.
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27. Peripheral Blood Smear Part 2
Blood Smear Procedure
Fresh drops of blood are obtained, usually following dermal puncture
Wipe away the first drop of blood
Place 1 drop of blood about 1/2 inch from the frosted end of one glass slide
Hold the spreader slide at a 30 to 35o angle and spread blood toward the opposite end of the slide
The blood smear should be smooth in appearance with no irregularities, streaks, or holes and a feathered edge
The wedge method is the most common technique for making blood smears
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28. Apply Your Knowledge
Blood smears are used to diagnose which of the following diseases?
Malaria
Leukemia
Anemia
All of the above
None of the above
Bravo!
ANSWER: D. All of the above
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29. Point-of-Care Testing
Near-patient testing
Designed to reduce hospital cost and reduce turnaround time for blood test results
Instruments are portable, internally calibrated, and easy to use
Point-of-Care testing (POCT) is usually performed by phlebotomists, nurses, technicians, or medical assistants
Dermal puncture usually done because a small blood specimen is needed
Each instrument must be calibrated and requires quality control checks
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30. Point-of-Care Testing Part 2
Bicarbonate
Ionized calcium
Cholesterol
Blood ketones
Urine dipstick
PT and PTT
Chloride
Blood gases
Common Point-of-Care Tests
Glucose
Hemoglobin
Sodium
Potassium
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31. Point-of-Care Testing
Often requires blood from a dermal puncture.
POCT is done at various facilities.
If a clinical lab is not available, only CLIA waived testing can be performed.
CLIA waived tests are those that are cleared for home use.
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32. Point-of-Care Testing
Before the Test
Confirm test orders
Establish a procedure for patient ID
Give patient pre-test instructions
Collect specimens according to package directions
Label specimens appropriately
Never use expired kits
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33. Point-of-Care Testing
During the Test
Perform quality control testing
Correct any problems
Establish a policy for frequency of control testing
Carefully follow all test-timing recommendations
Interpret test results using product inserts as a guide
Record test results according to office policy
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34. Point-of-Care Testing
After the Test
Report results in a timely manner
Follow package insert for follow-up
Follow OSHA regulations for disposing of biohazardous waste
Participate in quality assurance assessment programs
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35. Apply Your Knowledge Part 7
Which of the following tests cannot be performed with POCT?
A. Glucose
B. Hemoglobin
C. Blood culture
D. Cholesterol
Good Job!
Answer: C. Blood cultures require a larger amount of blood and have to grow in culture media.
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36. Urine Specimens Used to evaluated substances found in urine
Consist of the physical component, chemical component, and the microscopic component
First morning urine is best used for testing
Chain of custody must be established for drug and alcohol testing
Should be refrigerated if not tested within 1 hour
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37. Urine Specimens Obtaining Urine Specimens For a Female: For a Male:
Separate the skin folds and wipe front to back three times
Keep the skin folds apart and collect a midstream urine in the cup
Once ¾ full, place the lid on the cup
Label the specimen
For a Male:
Use 2 towelettes to clean the head of the penis
If uncircumcised, keep foreskin retracted while patient urinates and collect a midstream urine in the cup
Once ¾ full, place the lid on the cup
Label the specimen
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38. Urine Specimens Urine Specimen Testing The Urine Dipstick
Uses reagent strips to test urine for a number of substances
Keep strips in a cool dry area
Never remove them from the container until ready to test
Examine the strips for discoloration
Check the expiration date
Always run a control sample when opening a new supply
Write the date opened
Follow the manufacturer’s directions for accurate results
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39. Apply Your Knowledge Which is the best urine specimen for testing?
ANSWER: The first morning urine
GREAT!
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40. Centrifuge Operation Centrifuge Operation
Centrifuging is the spinning down or separating of cells from the liquid portion of the blood.
Specimens also require aliquoting (dividing specimens into separate containers)
Aliquoting for most laboratory tests requiring serum or plasma must be done within two hours or less
Test results could be altered
If the RBCs are left in contact with the serum, the glucose level would be decreased
What happens if the specimen is not separated in time?
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41. Centrifuge Operation Part 2
Types of Centrifuges
Refrigerated
Floor models
Tabletop models
The speed of rotation and the radius of the rotor head determine the relative centrifugal force of a centrifuge, which is expressed in gravity (g).
Most laboratory specimens are centrifuged at 750 to 1000 g for 15 minutes
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42. Centrifuge Operation Part 3
Place tubes of equal size and volume directly across from each other so the machine will be balanced
Never open the lid until the centrifuge machine has come to a complete stop
After centrifugation, always be sure the transfer tube is properly labeled before you begin to aliquot a specimen
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43. Apply Your Knowledge
Within what time frame should specimens be centrifuged for tests that require serum or plasma?
ANSWER: Within 2 hours
GREAT!
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44. CHAPTER SUMMARY
Specimens may require special handling or collection in order to be accurate.
Blood cultures must be drawn under strict aseptic technique to prevent false results.
When using a Unopette, collect the specimen with the pipette and mix the specimen carefully.
To perform a microhematocrit, obtain blood in a capillary tube and centrifuge. The separated cells can be used to determine the patient’s hematocrit.
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45. CHAPTER SUMMARY (cont’d)
Peripheral blood smears require a drop of blood on a slide. The blood is spread across the slide using a spreader slide.
Point-of-care tests are obtained and tested immediately at the point of care or near the patient. CLIA waived tests pose a minimal risk to the patient.
A urine specimen can be collected and a reagent strip may be used to determine the concentration of a substance.
When using a centrifuge, make sure the load is balanced.
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46. END OF CHAPTER 5
Choose a job you love and you will never have to work a day in your life.
-- Confucius
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