Lesson 2.

Specimen Handling and Processing

Blood Specimens
Criteria for Recollection/Rejection of Sample
- Goal of phlebotomist: provide acceptable samples for laboratory testing
- If the criteria is not followed, the sample must be rejected and recollected
1) Labels must have the patient’s complete name/hospital number/unique identifier
▪ Mislabeling
a) Mortal sin of a medtech
2) Sample must have on the label the name of the test to be performed
▪ Especially for body fluids
3) Sample must be collected within the time limits requested
▪ Follows a TAT or STAT
4) Urine samples must have the label on the container, not on the lid
5) Appropriate anticoagulant
6) Tubes may not be used beyond the expiration date
7) Certain samples may not be exposed to light
▪ Analytes that can be damaged when exposed to light:
a) Bilirubin c) Vitamin D
b) Vitamin C d) Vitamin E
▪ Wrap the tube with aluminum foil or use amber tubes
8) Delays in centrifuging or processing a sample will invalidate the results
▪ If not centrifuged right away, sample is degraded
9) Anticoagulated blood samples must be free from clots
10) Blood samples drawn above an IV line are unacceptable
▪ Last choice: antecubital fossa
11) Sample must be recollected if the results are not consistent with previous results on the patient
▪ If not consistent, sample must be reprocessed or recollected
Whole Blood
- Do:
1) Maintain at room temperature (15-30°C)
- Do not:
1) Freeze
2) Centrifuge
3) Place in direct contact with cool packs or ice
- Storage: ref temp (48 hours at 2-10°C)
o This maintains sample integrity before disposal
- Tests:
1) CBC 3) ESR
2) Reticulocyte count
Plasma/Serum
- Do:
1) Allow sample to clot in an upright position for 15-30 minutes
2) Centrifuge for at least 2200-2500 RPM
3) Separate using a pipette
- Storage:
1) Do not include the RBC
2) Ref temp (48 hours)
3) Frozen temp (1 year at below 20°C)
- Tests:
1) Crossmatching 3) Chemistry tests
2) Coagulation tests 4) Serology tests
Non-Blood Specimens
Body Fluids
- Container: glass and sterile
- Centrifugation: 15 minutes at 2200-2500 RPM
o Do not centrifuge for hematology or bacteriology tests
- Storage:
1) Room temp (7 days)
2) Refrigerated (7 days)
3) Frozen (28 days)
- Tests:
1) Glucose 3) EBC and RBC count
2) Total protein 4) Differential count
Body Fluid Source Collection Volume
Cerebrospinal fluid (CSF) Spinal space Lumbar puncture > 0.5 mL (clear)
(Do not accept RC count > 150)
Pleural fluid Lungs Thoracentesis Minimum of 50 mL
Peritoneal fluid Abdominal area Paracentesis Minimum of 50 mL or
> 500 mL
Pericardial fluid Surrounding the heart Pericardiocentesis Minimum of 10 mL

Urine
- Collection: mid-stream, clean catch
- Volume: 30-60 mL
- Container: screwcap lid
- Processing: within 2 hours
Physical Examination Color, Clarity
Chemical Examination SG, pH, Glucose, Protein
Microscopic Examination WBC, RBC, Bacteria, Epithelial cells

Stool
- Collection: freshly collected
o Should not contain urine, oil, tissue, and other contaminants
- Volume: 0.5-1.0 gram or pea-sized
- Processing: within 2 hours
Physical Examination Color, Consistency
Microscopic Examination WBC, RBC, Parasite, Yeast

Sputum
- Collection: deep respiratory secretions, not saliva
- Time: early in the morning, rinse mouth with water
- Volume: about 1 teaspoon or 5 mL
- Processing: immediately
- Tests:
1) Microbiology
i. Gram stain iii. AFB
ii. Sputum culture
Semen
- Collection: collected on-site or in the house and delivered to lab within 60 minutes
o No sex and masturbations 2-5 days before sample collection
- Container: sterile wide-mouth container
- Temperature: close to body temp and must not be refrigerated
- Processing: within 2 hours
- Tests:
1) WBC 3) Sperm count, movement, and shape
2) Volume
Oropharyngeal (OP) and Nasopharyngeal (NP) Swabs
- Collection:
1) OP: posterior pharynx and tonsillar areas
2) NP: nares parallel to the palate
- Optimal timing: within 3 days and no later than 7 days
- Swab type:
1) Sterile dacron
2) Rayon swabs with plastic shafts
- Handling: immediately into sterile vial with 2 mL of viral transport media w/out antibiotics
Nasopharyngeal (NP) Wash/Aspirate
- Collection: insert tube into the nostril parallel to the palate
- Optimal timing: within 3 days and no later than 7 days
- Handling: sterile vial at 4°C

Recommended Clinical Specimens in the Setting of an Unexplained Respiratory

Disease
Outpatients
1. Upper respiratory
a. NP and OP swabs
b. NP wash/aspirate
2. Lower respiratory
a. Sputum
3. Blood
a. Serum: acute (at onset), convalescent (3-6 weeks post onset)
b. Blood (plasma)
4. Urine
5. Stool
Inpatients
1. Lower respiratory
a. Brachoalveolar lavage c. Pleural fluid
b. Tracheal aspirate d. Sputum
2. Upper respiratory
a. NP and OP swabs
b. NP wash/aspirate
3. Blood
a. Serum: acute (at onset), convalescent (3-6 weeks post onset)
b. Whole blood (plasma)
4. Tissue
5. Urine
6. Stool
Fatal Cases
1. Tissue
a. Fixed tissue from all major organs
i. Lung v. Brain
ii. Heart vi. Kidney
iii. Spleen vii. Adrenals
iv. Liver
b. Non-fixed tissue from lung and upper airways
i. Trachea
ii. Bronchus
2. Lower respiratory
a. Brochoalveolar lavage c. Pleural fluid
b. Tracheal aspirate d. Sputum
3. Blood
a. Serum
b. Blood (plasma)
4. Deep lung swab for bacterial culture