Significance 

There are a variety of microorganisms found in the water of heatlhcare facilities that are not generally pathogenic for healthy individuals, but may be pathogenic for the immunocompromised patient. The conditions associated with increased risk of acquiring an infection from a waterborne pathogen include; hematological malignancy, immunosuppression, transplantation and burns. These infections can result in significant morbidity and mortality. The waterborne pathogen panel is designed for use in healthcare facilities to evaluate the potential for exposure to four waterborne pathogens: Legionella, Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter species.

Test Method 

Legionella spp.
Filter concentration method using BCYE, DGVP and CCVC media, incubation 36.5° + 0.5° C for 7 to 10 days.  Direct plating is also included in the event of high concentrations of the organism.  Heat treatment may be utilized on samples with bacterial overgrowth. Modified version of the ISO Standard 11731:1998, Water Quality-Detection and enumeration of Legionella, and ISO Standard 11731:2004, Water Quality-Detection and Enumeration of Legionella Part 2: Direct Membrane Filtration Method for Waters With low Bacterial Counts. SPL is accredited by the American Association for Laboratory Accreditation (A2LA) to perform this test.

Pseudomonas aeruginosa
Filter concentration method using M-PA-C selective media, incubation 41.5° + 0.5° C for 48 to 72 hrs.  Modification of the ASTM International Standard Test Method for the Isolation and Enumeration of Pseudomonas aeruginosa from water [D 5246-92 (Reapproved 2004)] with the addition of direct plating in the event of high concentrations of the organism. SPL is accredited by the American Association for Laboratory Accreditation (A2LA) to perform this test 

Stenotrophomonas maltophilia
Filter concentration method using Trypticase Soy Agar with imipenem selective media, incubation 36.5° + 0.5° C for 48 to 72 hrs.  Direct plating is also included in the event of high concentrations of the organism. Modification of the ASTM International Standard Test Method for the Isolation and Enumeration of Pseudomonas aeruginosa from water [D 5246-92 (Reapproved 2004)]

Acinetobacter species
Filter concentration method using MacConkey agar, incubation 36.5° + 0.5° C for 48 to 72 hrs.  Direct plating is also included in the event of high concentrations of the organism. Modification of the ASTM International Standard Test Method for the Isolation and Enumeration of Pseudomonas aeruginosa from water [D 5246-92 (Re-approved 2004)]

Turn Around Time

2–10 days

Test Code

100

Sampling

This panel includes culture for four waterborne pathogens: 1) Legionella spp. 2) Stenotrophomonas maltophilia  3) Acinetobacter spp.  4) Pseudomonas aeruginosa. The sample volume is 500 ml and should be collected as follows:

  1. Complete the Chain of Custody form
  2. Use waterproof pen to label bottle with sample location, description, and date.
  3. Turn on hot water and immediately fill the bottle (500ml).  Close bottle and invert to mix neutralizer.
  4. Place water sample in shipping pouch with absorbent material and close pouch using zip lock seal.

Sample Transport

  1. Ship the same day samples are collected for receipt in laboratory within 30 hours of collection. The time from sample collection to analysis should not exceed 30 minutes.
  2. Seal thermal pouch and place in box.
  3. Place completed Chain of Custody form in plastic document holder and pack in box outside of thermal shipping pouch.
  4. Ship overnight Monday–Thursday only to: Special Pathogens Laboratory, 1401 Forbes Avenue, Suite 209, Pittsburgh, PA 15219

Questions? Call 412-281-5335