Specimen Collection
& Handling Requirements

Specimen Labeling Requirements

All specimens submitted must include 2 patient identifiers (see below), and the collection date & time.

  • First and last name (must match requisition form)
  • Date of Birth
  • Other facility/client ID

Note that the same two identifiers must be on the requisition form as well.

TWO IDENTIFIERS are required for patient and specimen identification. This laboratory policy is based on the requirements for the College of American Pathologists (CAP), our accrediting body.

Preferred identification is a wristband with patient name and date of birth.

A location (i.e. room number) is not an acceptable identifier.

For patients who are capable of active communication and those wearing arm bands, they will be asked to state their full name and date of birth. This will be compared to the information documented on the test requisition form. If the information corresponds with the information on the requisition form, the specimen collection will proceed. If the information received does not correspond with the information on the requisition form, nursing staff will be sought to confirm the identity of the patient.

For patients who are not able to personally participate in the confirmation process and who are not wearing armbands, nursing staff will be sought to provide the identity of the patient. The name of the nurse providing the identity of the patient will be documented.

NICL Laboratories reserves the right to defer specimen collection if there is reasonable doubt in the proper identification of the patient or lack of support in attempting to confirm the identity of a patient.

General Specimen Collection Procedure

  1. Prepare test requisition with required patient information as outlined on page 7 of this manual. Date on the form should be the date of specimen collection, not the date the order was received. Make sure the test requested is indicated.
  2. When ordering a urine culture, be sure to indicate the method of collection.
  3. Identify the patient and explain procedure.
  4. Wash hands before and after specimen collection.
  5. Wear gloves during specimen collection.
  6. Handle specimen containers carefully – touch only the outside of container to avoid contamination.
  7. Collect specimen. (Refer to specific procedures as outlined on following pages)
  8. Replace caps securely to prevent leakage.
  9. Label specimen with patient’s full name and date of birth. Add source of culture and date & time specimen was collected. Culturettes must be labeled on the plastic tube, not the outer packaging.
  10. Fold the top copy (original) of the test requisition in half widthwise (top to bottom) with the patient’s name facing out. Retain the back copy for your files.
  11. The specimen bag has two pouches. Place the specimen(s) in the rear pouch and the test requisition in the front pouch.
  12. Seal the specimen bag.
  13. Refrigerate all specimen containers immediately after collection. Do not refrigerate culturettes. Keep these at room temperature before and after specimen collection.

Specimen Rejection Criteria

  1. Improperly collected specimen (improper specimen container, improper temperature).
  2. Unlabeled or mislabeled specimens.
  3. Containers that are not properly sealed (e.g. a leaky container, a swab not in a transport container.
  4. Insufficient specimen quantity.
  5. Specimens submitted with a needle (e.g. body fluid in a syringe).
  6. Formed stool specimens for Clostridium Difficile testing.

Specimen Refrigerator

Temperature Range

Refrigerator 2 to 8°C (35.6 – 46.4°F)
Freezer -15 to -3°C (5 – 26°F)

  1. A specimen refrigerator is provided by NICL Laboratories for the sole purpose of storing specimens until picked up by a laboratory employee.
  2. No other items (medication, food, etc.) should be placed in this refrigerator.
  3. Refrigerator temperature should be checked daily by a facility employee.

Remember to Review General Specimen Collection Procedures

Urine Specimen Collection & Handling

Routine Urinalysis

The optimal Urinalysis specimen is the first morning void or urine that has been in the bladder for at least four hours. Peri-care prior to specimen collection helps to prevent contamination.

  1. Remove blue cap from the specimen cup – place on a clean surface with the inside of the cup facing up.
  2. Have resident urinate directly into the sterile cup.
  3. Place the cup on a clean, flat surface and replace the blue cap securely. Carefully peel back the protective sticker to expose the rubber-covered cannula. This device is a sharp!
  4. Remove the C&S Preservative Tube (gray top) from the package. Push the C&S Preservative Tube (gray top) into the integrated transfer port. Hold in position until the flow of urine has stopped. Remove tube, and shake tube vigorously.
  5. Remove the UA Preservative Tube (cherry red/yellow top tube) from the package. Push the UA Preservative Tube (cherry red/yellow top tube) into the integrated transfer port. Hold in position until the flow of urine has stopped. Remove tube, and invert tube 8-10 times to mix the sample.
    25 ml of urine in the specimen cup is necessary for both Vacutainer tubes to fill.
  6. Place protective sticker back over the integrated transfer port.
  7. Label original specimen cup and both filled tubes with the patient full name, date of birth and date & time of collection.
  8. Fold the top copy (original) of the test requisition in half widthwise (top to bottom) with the patient’s name facing out. Retain the back copy for your files.
  9. The specimen bag has two pouches. Place the specimen(s) in the rear pouch and the test requisition in the front pouch.
  10. Seal the specimen bag. Refrigerate immediately after collection.

Reflex Urinalysis

The following criteria will be used to trigger a reflex to culture which will be performed automatically:

  • Nitrite Positive
  • Leukocyte Esterase at Moderate or Large
  • Presence of WBCs:
    Male >2-5/hpf
    Female >5-10/hpf

Urine Culture & Sensitivity

Cultures are appropriate when a patient is symptomatic of an infection. Using guidelines for the Clinical and Laboratory Standards Institute and The American Society of Microbiology, the laboratory will “work up” one or two urinary tract pathogens in colony counts of 10,000 col/mL or greater. This “work up” includes organism identification and antibiotic sensitivities. Organisms present in smaller numbers, as well as normal flora, will not to “worked up” in clean catch specimens.

Urine Collection Procedures

Equipment:

Urine Collection and Transport Kit, includes:

  • A 4.5 oz. screw-cap specimen cup with integral sampling device, sterile interior
  • Gray top C&S Preservative Plus Urine Tube
  • Red/Yellow Marbled Top Urinalysis Preservative Plus Conical Urine Tube
  • Castile Soap Towelettes
Procedure
  1. Use bedpan, commode, toilet or urinal dependent of functional level and cooperation.
  2. Remove cup with blue cap and towelette from kit. Unscrew blue cap and place on clean surface with inside of cap facing up. Open towelette.
  3. Cleanse with towelette as follows:
    Male
    • If not circumcised: hold foreskin back before cleansing
    • Wipe head of penis in a single motion with the first towelette
    • Repeat with the second towelette
    • Urinate a small amount into the toilet
    Female
    • Separate the labia
    • Wipe inner labial folds from front to back with a single motion with the first towelette
    • Keep labia separated and urinate a small amount into the toilet
  4. Place cup under stream and continue to urinate into cup and collect specimen. (Do not fill the collection container completely to the top.
  5. Finish voiding into the toilet.
  6. Place the cup on a clean, flat surface and replace the blue cap securely. Carefully peel back the protective sticker to expose the rubber-covered cannula. This device is a sharp!
  7. Remove the C&S Preservative Tube (gray top) from the package. Push the C&S Preservative Tube (gray top) into the integrated transfer port. Hold in position until the flow of urine has stopped. Remove tube, and shake tube vigorously.
  8. Remove the UA Preservative Tube (cherry red/yellow top tube) from the package. Push the UA Preservative Tube (cherry red/yellow top tube) into the integrated transfer port. Hold in position until the flow of urine has stopped. Remove tube, and invert tube 8-10 times to mix the sample.
    25 ml of urine in the specimen cup is necessary for both Vacutainer tubes to fill.
  9. Place protective sticker back over the integrated transfer port.
  10. Label original specimen cup and both filled tubes with the patient full name, date of birth and date & time of collection.
  11. Fold the top copy (original) of the test requisition in half widthwise (top to bottom) with the patient’s name facing out. Retain the back copy for your files.
  12. The specimen bag has two pouches. Place the specimen(s) in the rear pouch and the test requisition in the front pouch.
  13. Seal the specimen bag. Refrigerate immediately after collection.
Equipment:

Urine Collection and Transport Kit, includes:

  • A 4.5 oz. screw-cap specimen cup with integral sampling device, sterile interior
  • Gray top C&S Preservative Plus Urine Tube
  • Red/Yellow Marbled Top Urinalysis Preservative Plus Conical Urine Tube
  • Castile Soap Towelettes
Procedure
  1. Before preparing the sterile field, open the Urine Collection and Transport Kit and remove the blue-cap urine cup.
    Unscrew blue cap and place on clean surface with inside of cap facing up.
  2. Using sterile technique, clean the patient and insert the catheter.
  3. After the flow of urine has started, collect the mid-stream flow in the specimen cup. Do not collect the initial flow. Fill the cup halfway if possible.
  4. Place the cup on a clean, flat surface and replace the blue cap securely. Carefully peel back the protective sticker to expose the rubber-covered cannula. This device is a sharp!
  5. Remove the C&S Preservative Tube (gray top) from the package. Push the C&S Preservative Tube (gray top) into the integrated transfer port. Hold in position until the flow of urine has stopped. Remove tube, and shake tube vigorously.
  6. Remove the UA Preservative Tube (cherry red/yellow top tube) from the package. Push the UA Preservative Tube (cherry red/yellow top tube) into the integrated transfer port. Hold in position until the flow of urine has stopped. Remove tube, and invert tube 8-10 times to mix the sample.
    25 ml of urine in the specimen cup is necessary for both Vacutainer tubes to fill.
  7. Place protective sticker back over the integrated transfer port.
  8. Label original specimen cup and both filled tubes with the patient full name, date of birth and date & time of collection.
  9. Fold the top copy (original) of the test requisition in half widthwise (top to bottom) with the patient’s name facing out. Retain the back copy for your files.
  10. The specimen bag has two pouches. Place the specimen(s) in the rear pouch and the test requisition in the front pouch.
  11. Seal the specimen bag. Refrigerate immediately after collection.

If current catheter has been in place for more than 2 weeks, the catheter should be changed before obtaining the specimen. Allow sufficient time for the bladder to yield 25 mL of urine before new catheter insertion and follow instructions for “Straight Catheter”.

Equipment:

Urine Collection and Transport Kit, includes:

  • A 4.5 oz. screw-cap specimen cup with integral sampling device, sterile interior
  • Gray top C&S Preservative Plus Urine Tube
  • Red/Yellow Marbled Top Urinalysis Preservative Plus Conical Urine Tube
  • Castile Soap Towelettes
Procedure
  1. Clamp drainage tubing (below sampling port) for sufficient time to yield 25 mL urine.
  2. Open the Urine Collection and Transport Kit and remove the blue-cap urine cup. Unscrew blue cap and place on clean surface with inside of cap facing up.
  3. Disinfect sampling port with alcohol wipe and obtain 25 mL of urine. Expel urine into the cup.
  4. Place the cup on a clean, flat surface and replace the blue cap securely. Carefully peel back the protective sticker to expose the rubber-covered cannula. This device is a sharp!
  5. Remove the C&S Preservative Tube (gray top) from the package. Push the C&S Preservative Tube (gray top) into the integrated transfer port. Hold in position until the flow of urine has stopped. Remove tube, and shake tube vigorously.
  6. Remove the UA Preservative Tube (cherry red/yellow top tube) from the package. Push the UA Preservative Tube (cherry red/yellow top tube) into the integrated transfer port. Hold in position until the flow of urine has stopped. Remove tube, and invert tube 8-10 times to mix the sample.
    25 ml of urine in the specimen cup is necessary for both Vacutainer tubes to fill.
  7. Place protective sticker back over the integrated transfer port.
  8. Label original specimen cup and both filled tubes with the patient full name, date of birth and date & time of collection.
  9. Fold the top copy (original) of the test requisition in half widthwise (top to bottom) with the patient’s name facing out. Retain the back copy for your files.
  10. The specimen bag has two pouches. Place the specimen(s) in the rear pouch and the test requisition in the front pouch.
  11. Seal the specimen bag. Refrigerate immediately after collection.

24 Hour Urine Specimen Collection & Handling

Because proper collection and preservation of 24-hour urine specimens are essential for accurate test results, patient should be carefully instructed in the correct procedure.

NOTE: When a creatinine clearance is ordered, a blood creatinine level needs to be drawn within the 24 hour period of urine collection. This requires a separate requisition.

Important Note

There are several tests which require 24-hour urines and a preservative may need to be added by the laboratory based on which test has been ordered. (Preservative will be added when the urine specimen arrives at the laboratory).

  1. Call the laboratory to request a 24-hour urine container. Upon receipt of container, label specimen container with patient full name, date of birth, name of test ordered, date & time collection started and date & time collection ended.
  2. Identify patient and explain collection procedure.
  3. Communicate to all shifts the 24-hour period selected (often 7AM – 7AM) and that ALL urine is being saved. Follow our facility guidelines for HIPPA compliant communication.
  4. When the collection is scheduled to start, have the patient void. DISCARD THIS URINE. This is to be sure the bladder is empty. The test will continue until the next day at the same time. This first voiding IS NOT included in the specimen.
  5. Patient may void in bedpan, urinal or “hat” placed in toilet. Each time the patient voids, the urine must be poured into the 24-hour urine container immediately.
  6. 24-hour urine container must be kept refrigerated, or in ice if refrigeration is not possible.
  7. For the next 24-hours, SAVE ALL URINE voided by the patient.
  8. At the end of the 24-hour period have patient void. Include this urine in the 24-hour sample.
  9. Notify the laboratory when collection is complete. Labelled container with completed test requisition will be scheduled for pick up.

Remember to Review General Specimen Collection Procedures

Stool Specimen Collection & Handling

Specimen Requirements: 2 grams in a sterile container

(orange cap container)

  1. A sterile bedpan (if available) is best for collected stool specimens.
  2. Ask the patient not to urinate into the bedpan and not to put toilet paper into the bedpan.
  3. Using a wooden tongue depressor, transfer stool from the bedpan to the sterile container. For diarrhea, a second specimen container may be used as a scoop.
  4. Replace cap securely on container and label immediately with patient full name, date of birth and time & date of collection.
  5. If collecting a stool culture, transfer specimen to a Cary Blair transport swab (red cap). Hold specimen at room temperature until picked-up.
  6. All stool specimens collected in a regular sterile container must be refrigerated until picked up. Please keep container upright.

Clostridium Difficile Testing

  • The test for C. Difficile is a molecular amplification test, not a culture, and is designed for use with unformed stools only.
  • If a patient produces more than 3 non-formed stools within a 24 hour period (unless ilius obstruction is suspected), they should be tested for C. Difficile.
  • Collect diarrheal specimen as described above. The specimen should not be a formed stool. (The specimen should take the shape of the collection container)
  • Formed stools are rejected for testing as a patient may be colonized with toxin-producing strains of C. Difficile, which are not causing illness.
  • The specimen cannot be collected on a swab.
  • One specimen per patient per week is accepted.
  • Positive results are called to the facility.
  • Repeat testing following a positive test as a test of cure is not recommended since the patient may carry the toxigenic gene for months after clinical cure. It may be indicated, however, if the patient improves with therapy and then relapses after the completion of treatment.
  • Repeat testing following a negative test is not recommended within a week due to the high sensitivity of testing methods. Due to these guidelines, a specimen will be rejected within 7 days of a previous test, unless special arrangements are made in advance of the ordering process.
  • Treating a patient with a negative test does not prevent development of a C. Difficile infection, and may increase the subsequent risk of illness.

Occult Blood Collection & Handling

Specimen requirements: 2 thin fecal smears

(applied to the Occult Blood Card)

Occult Blood cards (without the developer) are provided only to those facilities requesting the laboratory to interpret the results and provide a report to the facility. Facilities which perform the test in-house should provide their own supplies.

  1. Complete the patient identification information on the front of the Occult Blood card.
  2. Open this front flap – “Open Tab”.
  3. A clean, dry bedpan should be used for stool collection.
  4. Ask the patient not to urinate into the bedpan and not to put toilet paper into the bedpan.
  5. Using the applicator stick that comes with the Occult Blood card, collect a small amount of stool.
  6. Apply a very thin smear inside Box A of the Occult Blood card.
  7. Reuse applicator stick to obtain a second sample from different part of stool. Apply a very thin
    smear inside Box B of the Occult Blood card.
  8. Allow smears to air dry and then close the cover.
  9. Fold the top copy (original) of the test requisition in half widthwise (top to bottom) with the patient’s name facing out. Retain the back copy for your files.
  10. The specimen bag has two pouches. Place the specimen(s) in the rear pouch and the test requisition in the front pouch.
  11. Seal the specimen bag. Do not refrigerate the card. It should be maintained at room temperature until picked up by the laboratory.

Remember to Review General Specimen Collection Procedures

Blood Collection & Handling

  • Since prolonged stasis can result in alteration of some chemical values, a tourniquet should be used for a minimum period of time (Recommended amount of time is less than one minute).
  • Blood should not be collected while intravenous solutions are being administered of collected in the same syringe used to inject these solutions.
  • Blood samples should be put into the tube appropriate for the test requested.
    ALL TUBES SHOULD BE GENTLY INVERTED SEVERAL TIMES IMMEDIATELY AFTER BEING COLLECTED. View Chart
  • Please notify the laboratory for specimen pick-up as soon as possible. Certain tests require specimen processing within a few hours of collection. Please contact the laboratory with questions about specimen stability.

Blood Collection Tube Order of Draw

To prevent contamination of the specimen with additives from the previous tube, collect in the following order:

  1. Blood Cultures
  2. Light Blue (sodium citrate)
  3. Red Top (no gel, serum)
  4. Gold Top or Red/Black Top (gel, serum)
  5. Green Top (sodium heparin)
  6. Lavender Top (EDTA)
  7. Gray Top (sodium fluoride)

Blood Collection Tubes

We supply our clients with Vacutainer® tubes for the collection of blood samples that are sent to us for testing. These tubes contain pre-measure vacuum and pre-measured additives; therefore it is very important that these tubes are filled to the “stated fill” volume. If you over-fill or under-fill the collection tube, there is the potential for incorrect test results being reported.

Most laboratory tests are performed on anticoagulated whole blood, plasma or serum. Collect blood specimens in the color-coded Vacutainer® tube indicated in the Test Color Guide.

Blood Collection Procedures

  1. Nursing staff should stop fluids running through all single, double, or triple lumen catheters for two minutes.
  2. Draw a discard volume of blood. Approximately 5ml is usually adequate.
  3. If a syringe will be used for sample collection, withdraw the blood slowly to reduce hemolysis. A 10ml syringe is appropriate for specimen collection.
  4. A tube holder may also be attached to the VAD and tubes filled in the correct order of draw after the appropriate discard.
  5. The nurse should then flush the line with about 10ml of saline or per facility flushing policy, and any infusing fluids resumed.
  6. Tubes must then be labeled at the bedside by the person who obtained the specimen and appropriate documentation completed.
  7. Facility policies and procedures should always be followed.

Check to be certain that an antimicrobial agent hasn’t been administered through the line in the last hour. It is essential to avoid drawing from lines within an hour of completion of antimicrobial agent administration, as the antimicrobial agent may be passed into the blood culture bottles and prohibit growth.

  1. Make sure you are wearing proper personal protection equipment (PPE), including gown and gloves.
  2. Gather the required equipment:

    (2-3) SPS yellow top tubes (have an extra on hand in case of problems)
    (4) alcohol preps
    (1) 3-5 cc syringe
    (2) 10 cc syringes or (1) 20 cc syringe
    (1) Blood Transfer Device (female luer adapter)

  3. Using two separate alcohol preps (70% alcohol), scrub catheter hub connection for 15 seconds with each wipe. Let air dry.
  4. Using the last two alcohol preps, carefully wipe the tops of the SPS yellow top tubes for 15 seconds each.
  5. Disconnect the tubing from the cap of the catheter and attach the small 3-5cc syringe to collect discard blood. The suggested amounts for adults are 3 ml and 0.2 ml for pediatric patients. This blood is used to wash the line, and is not used for culture. The entire syringe and contents can be discarded in a biohazard container.
  6. Using a new syringe, collect blood for culture through the hub. 10 ml is required for each of two (2) SPS tubes (the total required volume is 20 ml). When enough blood has been obtained for 2 tubes, detach the syringe from the catheter and reconnect the tubing to the cap of the catheter.
    Note: If you are using 10 cc syringes, you will be required to perform this procedure twice.
  7. Attach Blood Transfer Device to the syringe using sterile technique. Sterile technique requires that you do not touch the top of the alcohol cleansed tubes with anything prior to insertion into the Blood Transfer Device/Syringe assembly.
  8. Insert the first SPS tube into the Blood Transfer Device/Syringe assembly and allow blood to transfer to tube using the tubes vacuum. Do not depress the plunger of the syringe.
  9. When the appropriate number of tubes is filled, dispose of Syringe/Blood Transfer Device as one unit in a biohazard container.
  10. Mix SPS tubes gently 8 – 10 times to prevent clotting.
  11. Label each tube with the patient full name, date of birth, date & time of collection, and collection site.
  12. Fold the top copy (original) of the test requisition in half widthwise (top to bottom) with the patient’s name facing out. Retain the back copy for your files.
  13. The specimen bag has two pouches. Place the specimen(s) in the rear pouch and the test requisition in the front pouch.
  14. Seal the specimen bag. Do not refrigerate. It should be maintained at room temperature until picked up by the laboratory within 2 hours of collection.

Mixing Chart

  • Vacutainer® Tube Type
  • EDTA
  • Citrate
  • SST with gel
  • Serum
  • Sodium Heparin
  • Sodium Fluoride
  • Stopper Color
  • Lavender
  • Light Blue
  • Gold or Red/Black
  • Red
  • Green
  • Grey
  • Number of Inversions
  • 8-10
  • 3-4
  • 5
  • 5
  • 8-10
  • 8-10

Color Guide

Remember to Review General Specimen Collection Procedures

Microbiology Specimen Collection & Handling

Please use tight sealing sterile containers or tubes of transport medium that will maintain viability, prevent drying out of the specimen swab, and prevent overgrowth of nonpathogenic microorganisms. It is important to label the container with the patient name and source. The inoculated containers should show no sign of leakage.

Deliver specimens to the laboratory as soon as possible to ensure recovery of all clinically significant organisms. NICL Laboratories’ specimen collection supplies and procedures support sample viability for a minimum or 48 hours except where indicated.

General Considerations

  1. Whenever possible, specimens should be obtained before antibiotics or other antimicrobial agents have been administered.
  2. Clinical material should be collected in leak-proof specimen containers which are tightly sealed.
  3. Material should be collected where the suspected organism is most likely to be found and with as little external contamination as possible (this is particularly important for draining lesions).
  4. The stage of the disease is sometimes an important consideration in the successful isolation of the causative agent.
  5. Specimens should be of sufficient quantity to permit completion of all tests ordered.
  6. Provisions should be made for the prompt delivery of the specimen to the laboratory.
  7. Most clinical material can be held for several hours at 4°C in a refrigerator before culturing if it cannot be processed immediately. This is particularly true with the following specimen types: urine, sputum, and material on swabs taken from a variety of sources. DO NOT refrigerate stools (for enteric pathogen isolation) and body fluids such as CSF or blood.
  8. Specimens for Neisseria gonorrhoeae isolation MUST be submitted on appropriate isolation plates (Martin-Lewis or Neigon agar plates).
  9. All stools for Ova and Parasite exam require preservation in Proto-fix CLR preservative vial immediately after collection.
  10. Collect up to three (3) consecutive stools for culture and one (1) per day for O&P. In hospitalized patients, stool cultures/O&P exams should not be performed if the length of stay is greater than 3 days and the admitting diagnosis is gastroenteritis.
  11. For Mycobacterial (TB) culture, it is recommended to collect three (3) sputum specimens for acid-fast smears and culture. These three (3) samples should be collected at 24 hour intervals and should include at least one early morning specimen.

Viral and Chlamydia Tests

Viral and Chlamydia Cultures:

  • Viral Chlamydia Transport Media (M4RT) or equivalent. IMPORTANT: After sample collection, refrigerate culture specimens until pick-up.
  • DNA Probe: Viral Chlamydia Transport Media (M4RT). Refrigerate after sample collection. Specimen stable 7 days.

Collection Procedures

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