Blood Specimen Handling and Processing for Lab Testing

Handling and Processing of Blood Specimens for Laboratory Testing

Routine Handling

• Phlebotomist's roles:
  • Perform routine venipuncture
  • Prepare specimens for transport to the laboratory
  • Adhere to time limits for delivery of specimens

Steps Involved in Processing and Handling

• Patient identification
• Blood collection
• Choice of tubes
• Phlebotomist's responsibilities for each test

Mixing Tubes by Inversion

• Cap color and number of inversions:
  • Red (Glass): 0x
  • Light Blue: 3-4x
  • Red (Plastic), Gold/Tiger Top: 5x
  • Light Green, Green, Lavender, Pink, Gray, Tan, Yellow, Orange, Royal Blue: 8x

Transporting Specimens

• Avoid hemolysis, platelet activation, and coagulation
• Transport with stopper to:
  • Avoid contact with the content
  • Minimize agitation
  • Aid clot formation

Special Handling

• Body temperature: transport at near body temperature (37°C) for certain specimens
• Chilled specimens: slow down metabolic process for certain specimens
• Light-sensitive specimens: protect from light to avoid affecting test results

Blood Specimen Processing and Reasons for Specimen Rejection

• Specimen processing:
  • Identification
  • Logging
  • Sorting by department
  • Evaluation for specimen suitability
• Reasons for specimen rejection:
  • Inadequate identification
  • Inadequate volume
  • Hemolysis
  • Wrong tube for collection
  • Outdated tube
  • Improper handling/mixing
  • Contamination
  • Insufficient specimen (QNS)
  • Collection time incorrect
  • Specimen exposed to light
  • Procedure did not follow testing time limits
  • Delay or error in processing

Delivery Time Limits and Exceptions

• Delivery time limits:
  • Routine blood specimens: 45 minutes
  • Centrifugation: 1 hour
  • Hematology specimen (EDTA tube): should not be centrifuged
• Time limit exceptions:
  • STAT or emergency specimens
  • Specific exceptions for certain tests (e.g., blood smear, EDTA specimen for CBC, etc.)

Centrifugation

• Importance of leaving the stopper on the tube before centrifugation
• Balancing tubes in the centrifuge
• Preparation of plasma and serum specimens

Aliquot Preparation

• Definition of aliquot
• Preparation of aliquot for chemical analysis or testing

OSHA Act Requirements

• Required protective equipment for handling specimens
• Includes gloves, gown/coats, and masks

Handling and Processing of Non-Blood Specimens

Urine

• Most analyzed non-body fluid
• Inexpensive to test
• Importance of urine analysis in health monitoring and disease diagnosis

Urine Collection Methods

• Random
• Midstream
• Catheterized
• Suprapubic aspiration
• Pediatric

Urine Tests

• Routine urinalysis
• Urine culture and sensitivity
• Urine cytology studies
• Urine drug testing
• Urine glucose and ketone testing
• Urine pregnancy testing
• Other urine tests (e.g., electrophoresis, heavy metals, myoglobin clearance, etc.)

Types of Urine Specimens

• Random
• First morning/8-hour urine specimen
• Fasting
• Timed
• 24-hour urine collection
• Double-voided

Urine Collection Procedure

• Empty bladder upon waking up
• Affix label and write down time and date
• Start timing
• Discard line output at first hour
• Collect all urine for the next 24 hours
• Refrigerate specimen during collection period
• Collect urine prior to bowel movement
• Drink fluids as needed to avoid dehydration
• Take one last void at the end of 24-hour collection period
• Seal the container before transporting to the lab

Other Non-Blood Specimens

• Amniotic fluid
• Cerebrospinal fluid (CSF)
• Gastric fluid/gastric analysis
• Nasopharyngeal secretions
• Saliva
• Semen
• Serous fluid
• Sputum

Note: The above notes are a concise summary of the key points and focus on the most important information.### Sweat

• Used to analyze the chloride content of patients under 20 with symptoms of cystic fibrosis
• Sweat Chloride Test:
  • Iontophoresis (electrical stimulation) in the forearm or thigh to stimulate sweat production
  • Pilocarpine (sweat-stimulating drug) is used
  • Sweat is collected, weighed, and analyzed for chloride content
• Can also be used to detect illegal drug use by placing patches on the skin for an extended period

Pilocarpine Iontophoresis

• Test site is cleaned and dried, then a piece of pre-weighed filter paper is placed over the test site
• Sweat is collected for 30 minutes
• Filter paper is retrieved and weighed to determine the weight of sweat collected

Synovial Fluid

• Viscous fluid that lubricates movable joints
• Tested to determine conditions such as arthritis, gout, and other inflammatory conditions
• Collected in three tubes:
  1. EDTA/Heparin tube for cell counts, crystal identification, and smear preparation
  2. Sterile tube for culture and sensitivity
  3. Non-additive tube for macroscopic appearance, chemistry, immunology tests, and observing clot formation

Arthrocentesis

• A procedure to collect synovial fluid from a joint

Buccal (Cheek) Swabs

• Used to obtain loose cells inside the cheek for DNA analysis
• Less invasive and painless alternative to blood collection
• Sample is collected by placing the swab inside the cheek and gently massaging the area
• Swab is sent to the lab where DNA is extracted from the cells

Bone Marrow

• Examined to identify blood diseases
• Physician inserts a large-gauge needle into the sternum or iliac crest (hip bone) and aspirates 1-1.5 mL of specimen
• Hematology technologist makes special slides from the first marrow aspiration

Breath Samples

• C-Urea Breath Test (C-UBT):
  • Checks for the presence of Helicobacter pylori bacteria that damages the stomach lining
  • Patient drinks a special substance with synthetic urea, then breathes into a Mylar balloon at specified intervals
  • Breath specimens are analyzed for carbon-13, which confirms the presence of H. pylori
• Hydrogen Breath Test:
  • Helps detect carbohydrate digestion problems, such as lactose (milk sugar) and fructose (fruit sugar)
  • Detects bacterial overgrowth in the small intestine
  • Most accurate tolerance test
  • Patient must not take antibiotics two weeks prior to the test and should fast on the day of the test

Feces

• Collected to:
  • Determine gastrointestinal disorders
  • Analyze for the presence of intestinal ova and parasites
  • Culture and examine for the presence of pathogenic bacteria and viruses
  • Check fat and urobilinogen content
  • Test for the presence of occult blood
• Collected in a clean and wide-mouth container, sealed, and sent to the lab

Hair

• Used to analyze for trace and heavy metals
• Used to detect chronic drug abuse

Throat Swabs

• Mostly collected to aid in streptococcal infection detection
• Special kit contains a sterile polyester-tipped swab and a covered transport tube
• Tube contains transport medium

Tissue Specimen

• Usually collected using biopsy through which the tissue sample is removed for examination
• Phlebotomy should check the proper handling procedure, particularly if the specimen delivered is not immersed in a solution
• For genetic analysis, tissue samples should not be placed in formalin

Arterial Puncture

• Used to collect blood specimens for ABG analysis to manage cardiopulmonary disorders and maintain acid-base balance of the body
• Arterial blood is the ideal specimen for respiratory function evaluation due to the consistency of its composition and high oxygen content
• Composition of Arterial Blood:
  • Uniform throughout the body
  • Tests requiring arterial blood include blood gases (O2 and CO2), lactic acid, and ammonia

Arterial Puncture Equipment

• Collection Kits:
  • Syringes: preanticoagulated, plastic, 1-5 mL based on instrumentation and tests
  • Needles: 20- to 25-gauge, 5/8 to 1.5 inches
• Anticoagulant:
  • Heparin used to coat the plastic syringes, tested within 30 minutes
  • Glass syringes used for tests longer than 30 minutes, lubricated, and heparinized by collector
  • Tightly fitting cap for syringe Luer tip

Arterial Puncture Sites

• Sites:
  1. Radial Artery (most preferred site)
  2. Brachial Artery
  3. Femoral Artery
• Order of Preference: Radial > Brachial > Femoral
• Gauge of Needle Required: a. Brachial Artery: 18-20 gauge b. Radial Artery: 23-25 gauge### Adverse Events in Blood Donation
• Healthcare personnel must monitor donors for adverse effects during and after blood collection
• Donors should remain seated for a few minutes to ensure they are not suffering from dizziness

Donor Care Post Phlebotomy

• Venipuncture sites should be inspected
• Refreshments should be offered to donors before they leave the area

Donor Blood Processing

• Collected blood units are prepared and placed in proper containers for transport to the processing area

Donated Blood Labeling

• Information on the blood unit label must be double-checked, complete, and accurate

Requirements by Donation Type

Whole Blood Donation

• Donation frequency: 56 days
• Must be in good health and feeling well
• Must be at least 16 years old
• Must weigh at least 110lbs

Power Red Donation

• Donation frequency: Every 112 days, 3 times per year
• Must be in good health and feeling well
• Male donors: at least 17 years old, 5'1" tall, and 130lbs
• Female donors: at least 19 years old, 5'5" tall, and 150lbs

Platelet Donation

• Donation frequency: Every 7 days, up to 24 times per year
• Must be in good health and feeling well
• Must be at least 17 years old
• Must weigh at least 110lbs

Donation Intervals

• Wait at least 8 weeks between whole blood donations
• Wait at least 7 days between platelet donations
• Wait at least 16 weeks between Power Red donations

Who can donate?

• Women with menstruation (if they can)
• People who take vitamins (except those taking antibiotics)

Infectious Disease Tested for on Each Unit of Donor Blood

• Chagas disease (Trypanosoma cruzi)
• HIV 1 and 2
• Hepatitis B (Hepatitis B Virus)
• Hepatitis C (Hepatitis C Virus)
• Syphilis
• West Nile virus (West Nile Virus)
• Human T-cell lymphotropic virus (HTLV)

Minimum Requirements for Venipuncture Blood Donation

• BP monitors
• Scales
• Donor couches
• Chairs
• Beds
• Blood Collection Mixers
• Blood Bag Sealers
• Blood Transportation Boxes
• Blood Bank Refrigerators
• Furniture and equipment must be made of cleanable surfaces, kept clean, and disinfected with sodium hypochlorite bleach solution
• Closed collection system with a sterile blood collection bag containing anticoagulant with attached tube and needle

Donor Collection

• Cleansing
• One-step procedure
• Two-step procedure
• Needle sizes: usually 15-17 G
• Veins of choice: veins of the antecubital fossa
• Fist clenching: speed flow of blood
• Completion of Donation

One-Step Procedure

• Duration: about 1 minute
• Material: 2% chlorhexidine gluconate in 70% isopropyl alcohol
• Procedure:
  • Wash the site using firm gentle pressure
  • Make sure the whole skin site is covered and in contact with the disinfectant for at least 30 seconds
  • Allow the site to dry completely or for at least 30 seconds

Two-Step Procedure

• Duration: about 2 minutes
• Material: 70% isopropyl alcohol and tincture of iodine or chlorhexidine (2%)
• Procedure:
  • Follow the steps in the one-step procedure
  • Use either tincture of iodine or chlorhexidine (2%)
  • Start from the center, then move downward, and outward applying firm but gentle pressure
  • Contact should be about 30 seconds
  • Then, allow the area to dry completely or for about 30 seconds

Venipuncture

• After confirming that bleeding has stopped, phlebotomist should place a sterile gauze/bandage and instruct the donor to keep it on for 4 hours

Apheresis Blood Donation

• Involves the collection of red cells, platelets, or plasma
• Can be autologous or allogenic

Autologous Blood Donation

• Autologous: involving one individual as both donor and recipient
• Purpose: patients scheduled for surgery donate their own blood for own use to prevent transmission of blood-borne pathogens
• Qualifications:
  • Time between donations: as often as 72 hours with doctor's approval
  • Hemoglobin level: at least 11.0 g/dL
  • Hematocrit: 33%
  • Autologous units are only designated to autologous donor

Therapeutic Phlebotomy

• Removal of blood
• Formerly termed as bloodletting
• Used for polycythemia (increased RBCs) and hemochromatosis (increased iron levels)

Patient Instruction

• Collection instructions
• Verbal and written
• Prepared to answer questions
• Collection on site or at home
• Confirm with patients that they have understood the instructions