MICROBIOLOGY CULTURE INSTRUCTIONS                                                                                 Microbiology Programs

Inspect the fiberboard container carefully for all inclusions. After inventorying the contents of the container, store them in a refrigerator. These instructions are illustrated in the Program Guide online under Microbiology Instructions.

Special Safety Precautions - These specimens contain pathogens or potential pathogens and should be considered infectious and handled as though they are capable of transmitting disease. They should be handled and disposed of only by personnel trained to work with pathogenic bacteria. All laboratory precautions and safety measures appropriate to handling live cultures should be practiced when working with these specimens.

In addition to the Precautions section of the program guide, be especially careful to avoid aerosol creation, inhalation, ingestion or injection of bacteria. These specimens should be disposed of as hazardous waste.

Rehydration Instructions for Swab Samples – These specimens are stabilized viable microorganisms lyophilized directly to an inoculation swab. A Rehydration Fluid tube is provided for each swab. The Microbiology Rehydration Fluid is located with the rest of your samples and is not found inside the tube with the swabs. The Diluent vials are placed in a pouch labeled REHYDRATION FLUID. There are up to 5 equivalent vials, each containing approximately 0.5 mL of TSB. The vials are not specifically labeled for any sample or Microbiology program module. We recommend you label the rehydrating vial prior to use to avoid confusion.

a. Warm appropriate amount of media, one specimen swab and one vial of rehydrating fluid to room temperature (20-25ºC) for each specimen tested. Store the second swab and extra rehydrating fluid vials in the refrigerator in case repeat testing is necessary.

b. Remove swab from plastic tube and submerge the sample portion into the fluid. Allow approximately 10 seconds for the lyophilized specimen to liquify while mixing and swirling the swab gently.

c. Once the swab is saturated, inoculate your media directly with the swab. Return the swab to the rehydration fluid before inoculating each subsequent culture plate.

d. If you perform a gram stain as part of your identification procedure, you can now transfer a portion of the liquified specimen to a clean glass slide. Be sure to have inoculated all culture media prior to using the specimen swab to prepare your gram stain. NOTE: There will be a small amount of charcoal residue from the lyophilization process present on your gram stain slide. This should not interfere with your ability to determine the staining pattern of the organism.

e. Continue by following the procedure and methods used by your lab to identify the organism(s) present in these specimens.

For Uricult users or users of other tests that require a liquid sample.

a. Open each foil pouch at the tear slit and remove the swab from the foil pouch.

b. Submerge the swab portion only into the provided rehydration broth tube for 30 seconds to wet the swab.

c. Express as much material from the swab as possible by firmly pressing the head of the swab against the side of the rehydration broth tube.

d. Swirl the rehydration broth tube gently to ensure even mixing and then use the broth as you would a liquid sample.

Urine Colony Count (CC-) Samples – Each sample consists of lyophilized pellet in a labeled foil pouch and a labeled 99 mL bottle of dilution fluid.

a. Warm an appropriate amount of media, specimen pellet and 99 mL bottle of dilution fluid to room temperature (20-25ºC) for each specimen tested.

b. Remove the cap from the sample vial and open the flip-top on the dilution fluid bottle.

c. Empty the lyophilized pellet into the dilution fluid and recap the bottle securely. Be careful to avoid touching the pellet to avoid contamination.

d. Mix the contents of the bottle vigorously until the entire pellet has dissolved and the suspension is homogeneous in appearance.

e.The entire contents of the dilution bottle (which now contains the dissolved lyophilized pellet) simulates a urine sample.

  Proceed to test as you would a patient sample in your laboratory.

f. Continue by following the procedures and methods used by your lab to identify the organism(s) present in these specimens and/or perform colony count testing.

g. If you perform a gram stain as part of your identification procedure, you may transfer a small amount of the mixed solution from the first CC specimen to a clean glass slide. Be sure to use sterile technique when transferring the solution. NOTE: There will be a small amount of charcoal residue from the lyophilization process present on your glass slide. This should not interfere with your ability to determine the staining pattern of the organism.

h. Record your colony count results for only the first two CC specimens.

Reporting Results

1. Subject each specimen to your protocol for each source as described on your reporting form.
2. Code your answers using the Bacteriology Result Code List.  You must report proficiency results to the same level that you would report in the context of your specific laboratory practice. For example, if you identify organisms on your patient cultures to the species level, you must identify your PT cultures to the species level as well.
3. Participants reporting presumptive identifications by culture should use the specific Genus only result codes.  You MUST select a code that accurately reflects the limitations of your testing, or you may be flagged as incorrect.  When performing isolations only with selective media, report your results as you would a patient. Before selecting a result code, review all codes found on the Bacteriology Result Codes List, paying special attention to the source specific presumptive identification section.
4. For the BA- specimens, you must report all organisms present when specified on the reporting form. Failure to report all organisms as indicated will result in your lab starting with an automatic 50% score.
5. Antimicrobial susceptibility tests (ASTs) are to be performed on the first two urine culture specimens. AAB-MLE uses the current M100 and M45 CLSI standards for determining the appropriate antimicrobial agents based upon specimen source and organism isolated. Selection of an inappropriate antimicrobial will be scored as incorrect.  

ORGANISM RESULT CODES

Please review codes. Changes have been made to the organism codes.

Instructions for Handling INOCU-SWAB II™

Instructions for Handling INOCU-PELLETS