Specimen Collection & Handling

Specimen Collection & Preparation
Blood Collection
Blood Gas Analysis
Specimen Collection Tubes
Specimen Handling & Transportation
24-Hour Urine Collection
Random Urine Drug Screen Collection with Chain of Custody
Minimum Pediatric Volume of Whole Blood Required for Commonly Ordered Procedures
Ambient Specimens
Refrigerated Specimens
Frozen Specimens
Courier Services
Priority Codes



SPECIMEN COLLECTION INFORMATION

Specimen Collection and Preparation
Laboratory test results are dependent on the quality of the specimen submitted. It is important that all specimens and request slips be properly labeled in the presence of the patient. Specimens should be labeled with 2 patient identifiers (usually name and date of birth).  The collection date and time, and the origin (source) of the specimen, when applicable. If there is any doubt or question regarding the type of specimen that should be collected, it is imperative that Deaconess Hospital Laboratory be called to clarify the order and specimen requirements.

Blood Collection
Most laboratory tests are performed on anticoagulated whole blood, plasma or serum. In general, specimens should be refrigerated until placed in the courier box for transport to the laboratory. Please see our individual User’s Guide section for specific requirements.

  • Plasma: Draw a sufficient amount of blood with the indicated anticoagulant to yield the necessary plasma volume. Gently mix the blood collection tube by inverting 6-10 time immediately after draw. If required, separate plasma from cells by centrifugation within 20 minutes.  See "Specific Guideline for Coagulation" attached to Coagulation Assays in the alphabetical list.
  • Serum: Draw a sufficient amount of blood to yield the necessary serum volume. Gently invert tubes immediately following collection. Allow blood to clot at ambient temperature and then, separate serum from clot by centrifugation within 20-30 minutes of collection. Hemolyzed samples are not always acceptable.
  • Whole Blood: Draw a sufficient amount of blood with the indicated anticoagulant. Gently mix the blood collection tube by inverting 6-10 times immediately after draw.

Blood Gas Analysis

 Capillary

Absolute minimum collected in a special heparinized collection device is 125 mL.

Heparinized Syringe Collection

Absolute minimum in a special heparinized syringe is approximately .5m

 

Order of Draw

Lab Recommendations for Blood Collections from Lines

Drawing blood for routine testing below an IV:

  1. Turn fluid off
  2. Wait 2 minutes
  3. Discard 5 ml of blood
  4. Collect blood tubes according to the "Order of Blood Draw" instructions

Drawing blood for routine testing from central lines, implanted ports, and central venous catheters.

  1. Turn all fluids off
  2. Flush (all ports) with 5ml of normal saline (if there was fluid in the line/s).
  3. Wait 2 minutes
  4. Discard first 5 ml of blood
  5. Collect blood tubes according to the "Order of Blood Draw" instructions

Drawing blood for coagulation testing through any lines that have been flushed with heparin should be avoided if possible.  But if unavoidable, for collection from central lines, implanted ports, and central venous catheters:

  1. Turn all fluids off
  2. Flush (all ports) with 20 ml of normal saline (if there was fluid in the line/s)
  3. Wait 2 minutes
  4. Discard first 10 ml of blood
  5. Collect blood tubes according to the "Order of Blood Draw" instructions.

Processing Blood Specimens

Gold or Red Top Tubes
Allow blood to clot, usually 15-30 minutes, but no more than 1 hour except when a test specifies differently. Centrifuge for approximately 10-15 minutes until there is complete separation of cells from the serum

Prolonged centrifugation may cause hemolysis or evaporation. Short centrifugation will not separate serum from packed cells. Serum should not be in contact with cells for more than 45 minutes.

Do not re-spin the primary serum samples if the cells have been in contact with the serum for more than 2 hours. If a sample is over 2 hours old and the cells are remixed with the serum, aliquot the serum into a pour off tube and re-spin aliquot tube.

Dark Green Tops - usually only used on in-patient testing:

  • Must be 90% filled
  • Do not spin Ionized Ca or Quick Chem these tests usues whole blood and must be delivered to the lab immediately.

Light Green Top with separator (Lithium Heparin plasma):

  • Sample will not clot, otherwise, handle as stated above for gold or red top tube.

Lavender Top whole blood for CBC testing (Must be potassium EDTA)

  1. Visibly check for over-or-under fill, peeling back the labels if needed. Reject and recollect when:
    • Tube has less than 1/2 the state volume of tube's capacity
    • Tube contains more blood than the stated volume. Example: a capillary tube with blood above the 500 ul mark or a 2ml fill tube above the 2mL mark is over-filled.
  2. Gently rock the tubes by hand 2 or 3 times and inspect for gross clotting. If sample is questionable or a capillary collection, use wooden applicator sticks to rim sample. If clots are found, reject.

Blue Top vacutainers (3.2% buffered Na Citrate Plasma)

  1. Visibly check to ensure tube is adequately filled.  Tube must be no less that 90% of the total volume.  See the BD Vacutainer chart for the correct fill, if available. 
  2. Gently rotate/rock tubes by hand to check for visible clots before centrifugation.  If clotting has occurred, the sample must be rejected.
  3. Centrifuging: NOTE:
    Prefer all blue tops to be spun for 3 min at the “Normal Setting” in the
    Statspin for 3 minutes, if available.  Otherwise, spin at 17000 x g for 15 minutes.
    If the coagulation testing will not be completed within the whole blood sample stability time, aliquot the plasma into a plastic aliquot tube, removing only the top 2/3 of the platelet poor plasma from the specimen.
  4. Visually inspect centrifuged coag samples for hemolysis.  Check DH Test Directory for acceptability. 
  5. Visually inspect level of RBCs and plasma to determine if the HCT is above 55%.  If above, 55%, contact Hematology at 812-450-2482 or Gateway Hematology at 812-842-3170 for further instructions.
  6. Visually inspect the plasma for clotting.  Clotting will appear as fibrous strands with captured red cells “stringing” through the plasma or stuck to the side of top of the tube.  If clotting is present, the sample must be rejected.

Aliquoting Any Specimen

Some samples may need to be aliquoted due to not having enough original tubes or due to the test not going to be performed that day.  Or for samples to be sent to a reference lab.  Aliquot the samples by:
                       

  1. Print a computer label and place on the aliquot tube.
  2. Always check the accession number and the name of the aliquot with the original tube to ensure the same number and name before pipetting the sample into the aliquoted tube.
  3. Pipet the sample from the primary tube using a clean disposable pipette into a new disposable plastic, labeled aliquot tube.
  4. Write “Serum” or “Plasma with the anticoagulant” on the aliquot tube.  Ei. “Plasma-citrate” or Plasma-blue top”.
  5. Write the collection date/time/initials of collector on the aliquot tube unless these are available in the lab’s computer by viewing the accession #.

 

Specimen Collection Tubes

The following is a list of tubes referred to in the Laboratory User’s Guide specimen requirements:

Green-Top Tube (Sodium Heparin) and Green Top Tube (Lithium Heparin): This tube contains sodium or lithium heparin – used for drawing heparinized plasma or whole blood for special tests.

Note: After the tube has been filled with blood, immediately invert the tube several times in order to prevent coagulation.

Grey-Top Tube (Potassium Oxalate/Sodium Fluoride): This tube contains potassium oxalate as an anticoagulant and sodium fluoride as a preservative.

Note: After the tube has been filled with blood, immediately invert the tube several times in order to prevent coagulation.

Lavender-Top Tube (EDTA): This tube contains EDTA as an anticoagulant.

Note: After the tube has been filled with blood, immediately invert the tube several times in order to prevent coagulation.

Light Blue-Top Tube (Sodium Citrate): This tube contains sodium citrate as an anticoagulant – used for drawing blood for coagulation studies.

Note: It is imperative that the tube be completely filled. The ratio of blood to anticoagulant is critical for valid prothrombin time results. Immediately after draw, invert the tube 6-10 times in order to activate the anticoagulant.

Red-Top Tube: This tube is a VACUTAINER® containing no anticoagulant – used for drawing serum for selected tests.

Royal Blue-Top Tube: there are two types of royal blue-top Monoject® tubes – one with the anticoagulant EDTA and the other plain. These are used in drawing whole blood or serum for trace element analysis. Refer to the individual metals in the individual test listing to determine the necessary tube type.

Serum Separator (SST) Tube: this tube contains a clot activator and serum gel separator – used for various laboratory tests.

Note: Invert the tube to activate the clotting. Let stand for 20-30 minutes before centrifuging for 10 minutes. If frozen serum is required, pour off serum into plastic vial and freeze. Do not freeze VACUTAINER(S)®.

Special Collection Tubes: some tests require specific tube for proper analysis. Please contact Deaconess Hospital Laboratory prior to patient draw to obtain the correct tubes for metal analysis or other tests as identified in the individual test listing.

Yellow-Top Tube (ACD): This tube contains ACD – used for drawing whole blood for special tests.

Specimen Handling and Transportation

Serum plasma samples - Unless specific handling instructions are listed in the alphabetical listing (to follow), samples need to be processed within 2 hours of collection.

After centrifugation, samples should be refrigerated or frozen or testing performed within 8 hours unless other specific instructions are listed in the alphabetical listing.

Samples should be maintained at the same temperatures as required for sample stability whether samples are handled and processed in the building or whether samples are transported from other locations.

The correct sample type and special instructions are listed in the alphabetical test listing.

The collection of urine specimens is the responsibility of the nursing unit or client office except for those collected in the Laboratory Outpatient Area.

Specimens for routine urinalysis and cultures should be freshly collected specimens. All urines should be collected utilizing clean catch, sterile technique.

The nursing unit, attending physician or office staff will collect specimens for culture. See the Microbiology / Infectious Disease Specimens collection section of this User’s Guide.

24 Hour Urine Collection Instructions

Obtain an approved urine container from Deaconess Hospital Laboratory.

Do not urinate directly into the container. Urine should be collected in a clean container and then carefully poured into the 24-hour collection container.

              To collect a 24-hour urine specimen:

  1. Follow your physician’s directions regarding food, drink, or drugs before and during collection.
  2. Empty bladder completely on awakening in the morning and discard this urine specimen.
  3. Record date and time under “start” and begin the test. (EXAMPLE: 8/15/04 @ 7:00 A.M.). Keep the sample on ice or refrigerated unless otherwise instructed.
  4. All urine during the rest of the day and night for the next 24 hours must be poured into the container. (Note: If possible, keep container refrigerated during collection.)
  5. Make final collection the next morning at the same time and record under “finish.” (EXAMPLE: 8/16/04 @ 7:00 A.M.)
  6. Keep sample refrigerated unless otherwise instructed. Take the 24-hour specimen to your physician’s office or laboratory as soon as possible.

Random Urine Drug Screen Collection with Chain of Custody - Non Forensic Only

For use with a physician order requesting a drug screen with Chain of Custody.  Out-patient collection for chain of custody drug screen performed only at the Deaconess Hospital campus Out-patient area.

  1. Obtain a Chain of Custody form the Chemistry Section of the laboratory. Only use an approved Chain of Custody form as provided by Deaconess Regional Laboratory.
  2. Observe collection or put dye in toilet, turn off hot water and seal around bowl.
  3. Let patient choose a urine cup and instruct that you need a minimum of 50 cc.
  4. Take specimen from patient and note temperature and color on Chain of Custody form.
  5. Seal container and label with patient’s name and additional patient identification (date of birth or hospital number), date/time and collector’s initials.
  6. Complete the Chain of Custody form.
  7. Keep the top copy of the Chain of Custody form and place on the patient’s chart.
  8. Take the back copy of the Chain of Custody form and the urine sample to the laboratory.

Minimum Pediatric Volume of Whole Blood Required for Commonly Ordered Procedures

Hematology Procedures

CBC with diff - 500 ul - one lavender microtainer filled to the top

Chemistry Procedures

The following listing states the absolute minimum sample required for commonly ordered tests. This listing reflects just enough sample to perform the procedures once, and assumes no instrument problems or dilutions that require the sample to be repeated.

General Chemistry Assays

  Adult Newborns with Hemocrits around 75%
Basic Metabolic (Chem8) One full serum separator microtainer One and ½ full serum separator microtainer
Comprehensive Metabolic One and ½ full serum separator microtainer Two full serum separator microtainers
Single assays (such as those above) up to 8 assays One full serum separator microtainer One full serum separator microtainer
Osmolality One full serum separator microtainer Two full serum separator microtainer

 

 

 

 

Adult

Newborns

Carbamazepine

One full serum separator microtainer

One full serum separator microtainer

Dilantin (Phenytoin)

One full serum separator microtainer

One full serum separator microtainer

Phenobarbital

One full serum separator microtainer

One full serum separator microtainer

Gentamicin

One full serum separator microtainer

One full serum separator microtainer

Tobramycin

One full serum separator microtainer

One full serum separator microtainer

Valproic Acid

One full serum separator microtainer

One full serum separator microtainer

Vancomycin

One full serum separator microtainer

One full serum separator microtainer

 

SPECIMEN PACKAGING & COURIER SERVICE

Ambient Specimens
Small biohazard bags are designed to transport serum and urine specimens that do not require special temperature or handling.

Refrigerated Specimens
Couriers have the necessary equipment to transport refrigerated specimens to Deaconess Hospital Laboratory. Cooler bags are utilized to assure the specimen remains refrigerated. Place the specimen and accompanying request in a tightly sealed plastic biohazard bag.

Frozen Specimens
Our couriers have the necessary equipment to transport frozen specimens to Deaconess Hospital Laboratory. Dry ice is utilized to assure the specimen remains frozen. Send each frozen specimen in a vial not more than ¾ full and use plastic (not glass) vials. Place the specimen and accompanying request in a tightly sealed plastic biohazard bag.

Courier Services
Courier services are available for transporting specimens to Deaconess Hospital Laboratory from all locations in the city and many locations in the tri-state area. Regularly scheduled pick-ups can be arranged through Deaconess Regional Laboratory. Pick up frequency is determined by referral volume.

LABORATORY TEST ORDER ENTRY

Priority Codes

STAT – Need it drawn and tested ASAP

STAT2 – Need it drawn at a specific time and tested ASAP

TIMED – Need it drawn at a specific time, but tested routinely

ROUTINE – Routinely drawn and tested at the top of every hour

Remember : Use TIMED priority code for all tests requiring the specimen to be drawn at a specific time except:

  • AM rounds are always ordered with a ROUTINE priority code unless STAT results are required.
  • If STAT results are required for AM rounds (or any other specimen to be drawn at a certain time during the day), then the STAT2 priority code should be ordered. The TIMED priority code should rarely, if ever, be used for AM rounds.

Frequently Asked Questions

Q. Does ordering tests STAT speed up the process of downtime?

A. No, this actually delays the accessioning and testing of specimens during a downtime. Use priority codes appropriately as you would during normal operating procedures