General

Testing accuracy is inherently dependent upon careful attention to the collection technique and specimen preparation.

• Please do not send any specimens in glass containers unless specified in the test menu.

• Please do not leave needles on syringes, as this practice creates an unnecessary hazard to those transporting and receiving specimens and may invalidate results due to exposure to air.

• Specimens containing a preservative or anticoagulant should be mixed thoroughly.

• Avoid contamination with normal flora.

• If swabs are used, they must be placed in approved swab transport devices.

 

Anaerobic Cultures

The proper selection of specimens is extremely important for the laboratory confirmation of anaerobic infection. Since anaerobes are the predominant flora of the mouth, intestinal tract, and vaginal tract, specimens incorrectly taken from these sites may produce results that may be misleading. Specimens should be collected in a closed system (syringe, transtracheal aspirates, needle punctures, etc.) from deep sites, bypassing normal flora. Do NOT send syringe; transfer aspirate to Port-A-Cul vial (BBL 21608), A.C.T. tube (Remel 12401/12402), or other anaerobic transport tubes.

Small Tissues

Collect in anaerobic transport tube. Do not add any fluid. Replace stopper immediately.

Large Tissues

Collect in sterile jar or petri dish. Larger tissue pieces do not require anaerobic transport tubes, since anaerobiasis is maintained inside the specimen for a reasonable length of time. DO NOT ADD FLUID (saline, formalin, or alcohol).

Fluid Aspirates

Collect in a syringe without air; use a small syringe for a small amount of fluid. Expel aspirate into Port-A-Cul vial or other anaerobic transport tube.

Swabs

Return swab to the anaerobic transport tube which contains a reducing atmosphere and an indicator which is colorless in reducing conditions. Swabs are the least desirable specimens for anaerobic cultures. Avoid taking specimens with a swab if at all possible. Note: fungal and AFB cultures require additional samples in AEROBIC transport devices.

Blood Cultures

Avoid drawing blood from an indwelling catheter. If the specimen must be drawn through the catheter, indicate on the requisition. Not more than 3 specimens need be drawn in the initial 24-hour period.

Cannulas, IUD's, etc.

Submit in sterile containers. Note: Foley catheter tips are not useful for diagnosis of urinary tract infections.

Multiple Tests/Specimen Types

If two or more types of specimens (i.e., serum and plasma) are submitted on one patient, indicate the type of specimen and anticoagulant on each vial. Refer to the test menu for specimen handling specifications.

Plasma

Draw sufficient blood with the indicated anticoagulant to yield the required volume of plasma. Separate plasma as soon as possible, preferably by centrifugation. Transfer plasma to a plastic vial and indicate the anticoagulant used on the label. Avoid hemolysis, as some tests are invalidated by hemolysis.

Serum

Draw sufficient blood to yield the required volume of serum. Let specimens clot at room temperature unless otherwise specified, and separate serum within one hour, preferably by centrifugation. Transfer serum to a plastic vial unless otherwise specified. Avoid hemolysis, as some tests are invalidated by hemolysis.

Sputum

First morning specimen is best. Reduce irrelevant flora by rinsing mouth and/or cleansing tracheostomy site. Instruct patient on the need for sputum rather than saliva. Collect specimen in a sterile container. A good specimen will show purulent material. If the specimen is watery and almost clear, it is probably saliva and is therefore inadequate for culturing.

Stool

Special specimen containers are available for all testing on stool specimens. To permit expansion, the container should not be more than half full.

Swabs

Use only swabs approved for bacteriological examination and culturing. Q-tips and similar cotton tipped applicators may be bacteriostatic or bacteriocidal. 
Note: Please see our tests wiki for more information on Virology swab cultures.

Non-wound Specimens

May include throat, nasal, endocervical, urethral, rectal or vaginal sites. Firmly swab the area with a sterile, approved, cotton-tipped applicator. Return swab to plastic sleeve or insert swab deep into the transport medium and break the applicator just below the point where it was held.

Wound

Remove debris around the wound by gently wiping the area with cotton soaked in sterile saline or water (do not moisten the swab with fluid containing bacteriostatic substances or antibiotics). Obtain specimen from the most active site of the wound. Take care not to swab the normal skin area. Return swab to plastic sleeve or anaerobic transport device or insert swab deep into transport medium and break off applicator just below the point where it was held. On request form, clearly indicate site of wound.

Urine

Bacterial Cultures

It is of utmost importance to follow proper cleansing procedures for obtaining a clean catch or catheterized specimen. Submit in an approved transport device (BD urine transport which contains boric acid as a preservative) or refrigerate until transport. For Viral culture, clean catch urine without preservative is required. Morning voids have highest titers. See Appendix R [pdf]for Microbiology collection procedures.

Timed collections

A timed urine specimen should be collected for most assays. Urine collection containers are available on request. Refer to the test menu for each assay to determine the appropriate type and amount of preservative. For timed collections, patients should be given two containers and instructed to avoid direct contact with collection bottles or bags containing preservatives, especially hydrochloric acid. Patients should receive explicit instructions for obtaining a complete timed urine sample. Specimen containers should be labeled with the type of preservative added. See Appendix I [pdf], for appropriate urine preservatives.

Patient instructions for 24 hour urine collection:

• Avoid alcoholic beverages and vitamins for at least 24 hours BEFORE starting to collect urine and during the collection period. Do not discontinue medications unless instructed to do so by your physician, but inform the laboratory which medications you are taking.

• Start the timed collection period with the first morning void. DISCARD THIS VOIDING since the urine was formed prior to the collection period. Write the time on the container and requisition. COLLECT ALL SUBSEQUENT VOIDINGS UNTIL THE SAME TIME ON THE SECOND DAY, emptying the bladder and adding this last void to the collection container.

• All timed urine collection bottles should be REFRIGERATED during collection. Preservative may have been added to the container; this may be caustic. Please be careful not to spill any of the preservative. (If the patient has a catheter, the collection bag should be kept on ice between empties.)

• Inadequate preservative or loss of voided specimens are common errors encountered in the collection. As a guideline, START THE COLLECTION OVER if: a. any part of the specimen is lost. b. if diarrhea stool gets mixed with the urine.

• Measure and record the timed collection volume in the appropriate area on the test request form. Submit only the required aliquot of a well-mixed total collection. Refer to the test menu to determine the appropriate pH adjustment. Carefully following instructions for urine collections will avoid the necessity of recollecting specimens.

 

Random Collection

For random urines the patient should void into a clean container. The specimen should be labeled and refrigerated until pick-up. A clean-catch or midstream specimen is preferred. The patient should void a small amount of urine before collecting in a clean container.

Whole Blood

Use the anticoagulant indicated. Indicate the anticoagulant used on the label. Do not allow to freeze.

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Specimen Collection Tubes

The following is a list of tubes referred to in the Laboratory testing guide menu:

• ACD (Yellow-Top Tube)*:

  This tube contains acid citrate dextrose (ACD), which is used for special tests using whole blood. Do not substitute the yellow top (SPS) tube. SPS additive interferes with several assays that use ACD blood.

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• EDTA (Lavender-Top Tube)*:

  This tube contains EDTA as an anticoagulant and is used for most hematological procedures. Do not allow to freeze.

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• Lithium Heparin (Green-Top Tube)*:

  This tube is used for the collection of heparinized plasma or whole blood for special tests.

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• Potassium Oxalate/Sodium Fluoride (Grey-Top Tube)*:

  This tube contains Potassium Oxalate as an anticoagulant and Sodium Fluoride as a preservative, used to preserve glucose in whole blood and for some special chemistry tests.

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• PST (Lime Green Plasma Separator Tube)*:

  This tube contains heparin and plasma gel separator and is used for many different tests. After drawing specimen, invert the tube to mix before centrifuging for 10 mins. If frozen plasma is required, pour off plasma into plastic vial and freeze. This is used for STAT orders.

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• Serum (Red-Top Tube):

  This tube contains no anticoagulant. The tube is used for the collection of serum for selected chemistry tests and special immunohematology tests. An SST tube should never be substituted as the gel barrier can interfere with analysis.

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• Sodium Citrate (Light Blue-Top Tube)*:

  This tube contains sodium citrate as an anticoagulant, used for collection of blood for coagulation studies. This tube needs to be completely filled, the ratio of blood to anticoagulant is critical for valid coagulation results. After the tube has been filled with blood, immediately invert the tube gently several times to prevent coagulation. Reference ranges for coagulation studies are based on tube containing 3.2% buffered sodium citrate.

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• Sodium Heparin (Royal Blue-Top Tube)*:

  This tube is for lithium ratios only.

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• Special Tubes

  Other tubes for collection of special tests are available upon request. Please contact Reference Laboratory Services Services before collection.

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• SPS (Yellow Top Tube)*:

  This tube is for blood cultures (routine and AFB) for Microbiology.

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• SST Hemogard (5 mL) (Serum Separator Tube) - (Gold-Top Tube):

  The new preferred SST tube is a plastic 5mL Gold Hemogard (BD#367989) and replaces the plastic 8.5 mL red/black marble top serum separator tube currently in use (BD# 367991). This tube is a serum separator tube with clot activator and barrier gel to separate serum from the clot below. This tube is used for common chemistry panels such as the Basic and Comprehensive Metabolic Panels, Lipid Panel, and Hepatic Function Panel and any individual tests contained in these panels. This tube should not be used for drug analysis since some drugs can bind to the gel and yield incorrect results.

 

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

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Requisition Forms

Submission of requests for laboratory testing should be accompanied by the appropriate requisition form. Laboratory procedures are performed ONLY upon a physician's written request. All verbal requests for additional test requests to be added to a written order must be followed by written confirmation within 30 days. CLIA-88, and thus the Joint Commission and the College of American Pathologists (accrediting agencies) require that each requisition form for laboratory services must clearly identify:

• Patient name, sex, birth date, include unique I.D. number, lab reference number.

• Collection date and time

• Diagnosis Code

• Type of specimen submitted, indicate serial and timed collection

• Patient fasting conditions (if applicable)

• Interval and total volume if a timed urine collection

• Source of specimen if for Microbiology or Virology test(s), or other comments in "comments" box

• Authorization for laboratory work, include the name of the ordering physician and UPIN in addition to the name and address of the facility to which the results will be sent.

• Check test(s) requested. If the Department of Laboratory Medicine offers a procedure that does not appear on the request form, print plainly the full name of the test desired in the space marked "OTHER REQUESTS".

 

1. Complete a separate requisition for each patient.

2. Please provide all information on the left hand side of the requisition form.

3. Fold completed request form and place in the outside pocket of the Specimen Bag. If submitting more than one specimen per patient, and specimens need to be stored and transported at different temperatures, use separate bags and Test Request forms for each temperature type.

4. Special requests for testing should be noted on the requisition.

 

Labeling Specimens

• Please affix patient label vertically with the last name closer to the top end of the tube.

• Label or tag each specimen with at least patient name and/or unique I.D. number, and laboratory of origin, corresponding to the request form.

• Use dark, indelible ink. Be sure label or tag is secure.

• Use a separate request form for each specimen type or clearly mark specimen types on form.

 

All specimens must be properly labeled to ensure patient safety and prevent errors in patient diagnosis and treatment based on misidentified labels. Mislabeled/Unlabeled specimens will be rejected by UW Reference Laboratory Services Services.

Mislabeled specimens include:

• Specimens that are not labeled

• Specimens labeled with a patient name and/or patient identification number different from that on the accompanying laboratory request form.

• Specimens drawn from correct patient but labeled with wrong name and identification number.

• Specimens that have labels and requisitions that agree but have been drawn from the wrong patient.

 

Whenever a specimen is mislabeled Reference Laboratory Services area will notify the ordering location of the error and request a new specimen. Re-labeling will not be allowed except in the extremely rare instances where it may cause undue risk to the patient (e.g., CSF from lumbar puncture, tissues from a biopsy, or other similarly obtained irreplaceable sample). For these rare cases a clinical pathologist or designee will have to approve the request.

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Shipping Specimens

To avoid specimen rejection because of improper handling and degradation in transit, please follow the specimen requirement instructions in the test menu section of this guide. Any questions should be directed to Reference Laboratory Services (206) 520-4600 or (800) 713-5198. All specimens must be bagged for handling. The request forms must be in a separate compartment or bag.

Ambient Temperature Specimens

These specimens do not require special temperature conditions, but should not be allowed to freeze. However, safe specimen transport guidelines must be followed when specimens are submitted.

Clinical Specimens & Biological Substances

Any human or animal material, including but not limited to, excreta, secreta, blood and its components, tissue and tissue fluids, being shipped for diagnostic or investigational purposes must be packaged and shipped correctly. To determine the proper packaging, see Transport and Packaging of Specimens [52k pdf*] for details. All specimens prepared for transport to the University of Washington Medical Center or Harborview Medical Center should follow these general instructions. Please follow the appropriate directions whether you intend to use our courier, the bus, airfreight, or mail services (clients should follow any additional guidelines established by the U.S. Postal Service or commercial carrier they are using for safe transport of biological specimens). All specimens should be considered biohazards and handled accordingly.

NOTE: OSHA requires that all shipments containing clinical specimens be marked with a "Biohazard" label. Labeled bags for shipments sent to Reference Laboratory Services will be provided on request. Containers for shipping are also available through Reference Laboratory Services (206) 520-4600 or (800) 713-5198, upon request.

Frozen Specimens

Certain specimens must be shipped frozen to maintain stability of the analyte. Containers should be filled with dry ice to prevent samples from thawing in case of shipping delays. Request slip(s) and specimen(s) should be shipped in separate plastic bags. Specimens should be shipped in containers that have been tested and are known to keep samples frozen for at least 48 hours. Do not ship frozen specimens in glass containers unless specified under specimen requirements. Mark the outside of the container:

FROZEN MATERIAL - DO NOT THAW

 

Packaging and Transport

As of March 1, 2003 we would like all clients sending specimens to us to adopt a uniform specimen packaging procedure. In order to insure the safety of our couriers, and to make our processing workflow as streamlined and trouble-free as possible, we ask that you follow these guidelines:

• All specimens should be contained in sealed plastic specimen bags.

• The specimen bags should display the Biohazard logo

• Each specimen bag should contain enough absorbant material to contain all of the contents of specimen tubes in case of breakage.

• Each specimen bag should contain only ONE patient's specimen(s)

• Each specimen should be accompanied by the corresponding requisition sheet, folded and placed in the outside pocket of the specimen bag.

 

We will provide ziplock specimen bags upon request. To order bags, please call (206) 520-4600.

Refrigerated Specimens

For refrigerated specimens, use the same Styrofoam mailing container as is provided for frozen specimens. Request slip(s) and specimen(s) should be shipped in separate plastic bags. Place on a coolant pack and fill the vacant space with any type of packaging material. We request that coolant packs be shipped in plastic bags. Mark the outside of the container:

REFRIGERATED MATERIAL - DO NOT FREEZE

 

Radioactive Specimens

Specimens which contain radioisotopes will be accepted for testing only when prior arrangements have been made and the specimens are clearly labeled so as to identify the contaminating isotope, the dose and compound administered to the patient, and the date of administration. To minimize hazards, specimens which are very "hot", e.g. those taken immediately after ablative therapy, may be held until they are less radioactive or they may be analyzed by methods which are not routine in our laboratories. Such options will be discussed with you when you make arrangements to send the specimen.