Specimen Collection Guide
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 Online Testing Guide.
- 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.
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.
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.
Use the anticoagulant indicated. Indicate the anticoagulant used on the label. Do not allow to freeze.
The following is a list of tubes referred to in the Online Testing Guide.
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.
EDTA (Lavender Top Tube)*:This tube contains EDTA as an anticoagulant and is used for most hematological procedures. Do not allow to freeze.
EDTA (Pink Top Tube)*:This tube contains EDTA as an anticoagulant and is used for blood banking and transfusion service. This tube must be accompanied by requisition following specific patient identification procedures. Please see test guide or contact Transfusion Support Lab (TSL).
Lithium Heparin (Green Top Tube)*:This tube is used for the collection of heparinized plasma or whole blood for special tests.
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.
PPT (Pearl Top Tube)*:Plasma Preparation Tube (PPT) with K2EDTA and Gel for Separation, spray-coated K2EDTA Tubes are used for whole blood hematology determinations and immunohematology testing.
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 minutes. If frozen plasma is required, pour off plasma into plastic vial and freeze. This is used for STAT orders.
QuantiFERON (Gray, Green, Yellow, Red Top set of Tubes)**:QuantiFERON-TB Gold (QFT) comes in a set of four (4) tubes and is used to aid in the detection of Mycobacterium tuberculosis, the bacteria which causes tuberculosis (TB). Immediately after filling the tubes, shake them ten (10) times just firmly enough to make sure the entire inner surface of the tube is coated with blood. This will dissolve antigens on the tube walls.
RTS (Orange Top Tube)*:Rapid Serum Tubes contain a proprietary thrombin-based medical clotting agent and a polymer gel for serum separation. They are used for serum determinations in chemistry.
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.
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.
Sodium Heparin (Royal Blue Top Tube)*:This tube is for lithium ratios only.
Special TubesOther tubes for collection of special tests are available upon request. Please contact Client Support Services before collection.
SPS (Yellow Top Tube)*:This tube is for blood cultures (routine and AFB) for Microbiology.
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.
Streck Cell-Free DNC BCT (Brown Brick Top Tube)*:Cell-Free DNA BCT contains the anticoagulant K3EDTA and a cell preservative in a liquid medium. Cell-Free DNA BCT stabilizes cell-free plasma DNA as well as preserves cellular genomic DNA present in nucleated blood cells and circulating epithelial cells (tumor cells) found in whole blood. Tube must be accompanied with requisition specific to this test.
*Note: After the tube has been filled with blood, immediately invert the tube gently several times to prevent coagulation.
**Note: Immediately after filling the tubes, shake them ten (10) times just firmly enough to make sure the entire inner surface of the tube is coated with blood.
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 Online Testing Guide for specimen handling specifications.
DNA and RNA specimens submitted for testing must have been isolated using appropriate methods in a CLIA-certified laboratory. Requirements for minimum amounts, concentration, quality measures and storage are specified in the Online Testing Guide.
The proper selection of specimens is extremely important for the laboratory. While not always feasible, specimens should be collected before the administration of antimicrobial agents. Specimens should be collected in a manner which minimizes contamination with indigenous flora. When multiple tests are ordered, sufficient samples or volume should be submitted in order to provide for the ideal recovery of organisms. Specimens should be collected using appropriate collection devices and should be tightly sealed to prevent leaks. Specimen containers should be bagged and delivered to the laboratory in a timely manner under the appropriate transport temperatures for optimal recovery of microbial organisms.
Transport piece of tissue in an empty sterile container. Do not add fluid to the container. Larger pieces of tissue are ideal and maintain anaerobic conditions within the interior of the tissue.
Aspirate the fluid using a syringe. Remove the needle and replace it with a stopper. Alternately, collect the fluid in an empty sterile container. Do not add fluid to an ESwab as this will dilute the specimen and may compromise culture results.
ESwabs are recommended for transport and processing of specimens. Do not remove the liquid transport media in the ESwab as this is needed for processing the sample. Alternatively, samples may be collected using Amies gel transport swabs for culture, however they may not be suitable for all tests. Dry swabs are not acceptable for bacterial culture testing.
After swabbing the site to be tested, return swab to plastic sleeve or insert swab deep into transport medium and break off applicator just below the point where it was held or the scored mark on the shaft of the swab. On request form, clearly indicate site of the body the sample was collected from. Note: See below for more information on Virology swab collection.
Non-Wound Specimens (bacterial cultures)
May include throat, nasal, endocervical, urethral, rectal or vaginal sites.
Wound (bacterial cultures)
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.
Recommended 2 sets per 24-hour period. Each set contains one aerobic (Redox1) and one anaerobic (Redox2) bottles. Ideally add 10 mL of blood to each bottle. Note the original volume of media in the bottle before inoculating and add to a 10 mL mark above. Do not overfill bottles. The minimum volume of blood for an adult is 5 mL in each bottle. For infants the minimum volume is 0.5 mL in one aerobic bottle. Indicate the draw site for each set on the requisition. For UW Medicine only: If unable to draw directly into a blood culture bottle, draw into a SPS tube (not ACD) and the specimen will be transferred to bottles in the UW Microbiology laboratory.
For quantitative blood cultures blood must be drawn from the same site into a SPS tube in addition to both the aerobic and anaerobic bottles. Quantitative cultures will be rejected if the SPS tube is not collected.
For blood cultures for Acid Fast Bacillus (AFB) or for organisms that grow best on plated media (Histoplasma sp. or Bartonella sp.) the blood must be drawn into a 10 mL Wampole Isolator tube. The Isolator tube must be filled to the line on the side. If the tube is not filled completely it must, at a minimum, be half full otherwise the blood to anticoagulant ratio is not correct and the specimen will be rejected. In cases of short draws or pediatric patients it is recommended to use a pediatric 1.5 mL Isolator tube.
Miscellaneous Devices, Hardware or Catheter Tips
Transport in sterile containers as quickly as possible. Catheter tips specimens should be the segment near the skin and tip end and must not exceed 2 inches for a quantitative culture. Longer tip segments can be cultured but will not be given a colony count. Note: Foley catheter tips are not useful for diagnosis of urinary tract infections.
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.
Special specimen containers are available for all testing on stool specimens. Transport in Cary-Blair for bacterial culture and Enteric Pathogen PCR testing. For parasite exams transport in EcoFix vials. For fecal leukocytes or C. difficile testing transport fresh stool in a clean container. Formed stools will be rejected for C. difficile testing. Container lids should be tightly sealed to prevent leaking into the specimen bag.
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.
A timed urine specimen should be collected for most assays. Urine collection containers are available on request. Refer to the Online Testing Guide 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 Online Testing Guide to determine the appropriate pH adjustment. Carefully following instructions for urine collections will avoid the necessity of recollecting specimens.
Random Urine 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.
Gently insert mini-tipped flocked nasopharyngeal swab (swab on flexible plastic shaft) through the nostril and into the nasopharynx, reaching the posterior nasopharynx. Gently rotate swab, then remove, and break swab off into universal transport media (UTM). Make sure the swab is fully immersed in the UTM, and that the shaft is short enough to completely tighten the cap. Send to the laboratory as soon as possible. This specimen is appropriate for PCR or culture for many respiratory viruses, including RSV and Influenza A/B. Obtaining columnar epithelial cells will enhance reliability of viral detection. Maintain at 4° C before and during transport to the laboratory.
Sputum, BAL, Bronchial Washes and Tracheal Aspirates
These specimens should be placed in sterile containers. Maintain at 4° C before and during transport to the laboratory.
Rub regular sized flocked swab over posterior pharynx or both tonsillar fossae. In cases of mumps, swab the inflamed orifices of Stenson's ducts. Swabs should be vigorously rubbed over any open lesions in mouth or throat. Break or cut swab making sure that the swab shaft doesn't extend beyond the top of the vial, secure cap and check to see that the swab end is immersed in the universal transport media (UTM). Maintain at 4° C before and during transport to the laboratory.
Gently insert regular flocked swab into rectum enough to dirty it. Break swab off immediately into universal transport media. Maintain at 4° C before and during transport to the laboratory.
Conjunctiva (Eye) Swab
Evert the lower eyelid and gently rub the conjunctival surface with mini-tipped flocked swab. Swabbing pus collection in corner of eyes is not adequate. Break swab off immediately into universal transport media. Maintain at 4° C before and during transport to the laboratory.
Collect 5-10 grams of stool in clean container. Maintain at 4° C before and during transport to the laboratory.
Clean catch should be collected in sterile container. First morning voids usually contain the highest titers of virus. Maintain at 4° C before and during transport to the laboratory.
Cerebral Spinal Fluid (CSF)
Should be collected in sterile container. PCR is a much more sensitive method for diagnosing viral infections from CSF. 0.2 mL per PCR test ordered is needed. To culture CSF, a minimum of 0.5 mL of fluid ensures a complete viral work-up. Do not dilute in universal transport media (UTM). Maintain at 4° C before and during transport to the laboratory.
Miscellaneous Body Fluids
The general rule is that all body fluids should be collected in sterile containers, without transport media. Maintain at 4° C before and during transport to the laboratory.
Viruses can often be isolated from vesicular-pustular lesions. Macular lesions do not yield high titers of viruses. Vesicular lesions should be opened with a scalpel blade and vigorously swabbed with a regular or mini-tipped flocked swab or dacron-tipped swab. The swab should be placed in a vial of universal transport media (UTM), maintained at 4° C and transported to the laboratory as soon as possible. Lesion specimens collected in this manner provide material both for PCR or viral isolation. Swabs for PCR testing do not need to be placed in transport media, but specimens for culture must be transported in UTM.
Small pieces of tissue should be placed in universal transport media or sterile PBS. Do NOT add the flocked swab to the transport media. Large pieces may be transported in dry sterile container, as long as the specimen will be moist when it arrives in the laboratory. Hold specimen at 4° C and transport to the laboratory as soon as possible. Tissues for PCR testing do not need to be placed in transport media.
Bone marrow should be collected in an EDTA tube for either PCR or viral isolation. Heparin is inhibitory to PCR reactions and should be avoided. Specimens should be held and transported at room temperature.
Submission of samples 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 testing on specimens already submitted must be followed by written confirmation within 30 days. CLIA-88, the Joint Commission, and the College of American Pathologists (CAP) 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.
- 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".
Complete a separate requisition for each sample and please provide all information on request form. Fold completed request form and place in the outside pocket of the Specimen Bag. If submitting multiple specimens with different storage temperature requirements, use separate bags and requisitions for each sample. Note any special requests for testing on the requisition form.
When submitting specimens to the University of Washington Department of Laboratory Medicine, please complete a test order form and a supplemental testing form (if applicable) and send with the specimen(s).
For information, assistance, or for pre-addressed labels, call (206) 520-4600 or (800) 713-5198.
- Please affix patient label vertically with the last name closer to the top end of the tube.
- Label or tag each specimen in English with at least two of the following: patient name, date of birth 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.
- Please affix patient label vertically with the last name closer to the top end of the tube.
- Label or tag each specimen in English with at least two of the following: patient name, date of birth 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.
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.
- Specimens or requisitions with <2 matching patient identifiers.
Whenever a specimen is mislabeled, the ordering location will be notified of the error and requested to send a new specimen. Re-labeling will not be allowed except in rare instances where recollection may cause undue risk to the patient (e.g., CSF from lumbar puncture, tissues from a biopsy, or other similarly obtained irreplaceable samples). For these rare cases a clinical pathologist or designee will have to approve the request.
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 Client Support 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 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 Client Support Services will be provided on request. Containers for shipping are also available through Client Support Services (206) 520-4600 or (800) 713-5198, upon request.
Certain specimens must be shipped frozen to maintain stability of the analyte. Shipping containers should be filled with dry ice to prevent samples from thawing in case of shipping delays. Requisitions and specimens should be shipped in separate plastic bags. Specimens should be shipped in containers that have been tested, safe for use with dry ice, 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 with label: FROZEN MATERIAL - DO NOT THAW.
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 absorbent material to contain all of the contents of specimen tubes in case of breakage.
- Each specimen bag should contain only ONE patient's specimen.
- Each specimen should be accompanied by the corresponding requisition sheet, folded and placed in the outside pocket of the specimen bag.
Each patient's specimen(s) and requisition sheet must be contained in a single, separate specimen bag. We cannot accept specimens and paperwork from multiple patients if they are gathered together in a single bag.
For refrigerated specimens, use the same Styrofoam mailing container as is provided for frozen specimens. Requisitions and specimens 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.
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.
Courier service is provided at no charge* in the areas near Auburn, Bellevue, Bellingham, Bothell, Edmonds, Everett, Kent, Kirkland, Mt. Vernon, Olympia, Renton, Seattle, and Tacoma. The courier route includes bus and air freight depots. Stops are made at Sea-Tac Airport as needed. To schedule a pickup, please telephone us. Stops are made at the Greyhound Bus Terminal in the morning and afternoon, Monday through Friday. See Transport and Packaging of Specimens.
There is no STAT courier service.
All fees incurred for specimen transport other than via our courier service are the responsibility of the patient/sending location.
*Please contact Client Support Services office at 206.520.4600 or 1 800 713-5198 if you intend to use our courier service for non-departmental delivery.
The address you select for packages will vary according to the urgency of the package and the time it is sent. Routine specimens for ANY departmental laboratory (including Virology, Toxicology, Immunology, Microbiology, Hematopathology etc.) may be sent to the address below:
- UW Medicine
- Department of Laboratory Medicine
- 1959 NE Pacific Street, room NW220
- Seattle, WA 98195
- Tel: (206) 520-4600 or 1 (800) 713-5198
For packages sent by BUS or PLANE, and needing to be picked up, please add:
- Call on Arrival (206) 598-6224
It is recommended that you notify the Client Support Services Office (206) 520-4600 or (800) 713-5198 during business hours, 8 AM-5 PM Monday through Friday, prior to sending packages by bus or plane. At times outside of these hours, please call the laboratory directly at (206) 598-6224.
Because some of our laboratories are in separate locations, STAT toxicology specimens may be shipped directly to Harborview Medical Center. Please call (206) 744-3451 before shipping STAT specimens.
- Harborview Medical Center
- Clinical Laboratory, Room GWH47
- 325 9th Avenue
- Seattle, WA 98104
- Call on Arrival - 206.731-3451 RUSH
When sending to this address, please ensure all of the following is done:
- Do not send routine specimens to this address.
- Mark STAT or RUSH on the outside of the package as well as on the request form.
- Include the telephone number on the outside of the package.
Correspondence concerning billing may be sent via First Class Mail to:
- Client Support Services
- Department of Laboratory Medicine
- P.O. Box 15290
- Seattle, WA 98115-0246
Please do not send specimens to this address. For information, assistance, or for pre-addressed labels, call (206) 520-4600 or (800) 713-5198. For emergency assistance evenings and weekends, call the Laboratory Medicine resident on call at the UWMC Paging Operator, (206) 598-6190.
For information, assistance, or for pre-addressed labels, call 206.520.4600 or (800) 713-5198.
Last updated: 10/29/2018