ACCEPTABLE SPECIMENS:
Lung biogpsy, pleural fluid, bronchial washings, bronchial brushings, transtracheal aspirates, expectorated sputum, or blood.

Causes for Rejction:
1. Specimen not labeled with patient's name, MRN, date and time of collection, and collector's initials.

Collect specimen as follows:
BRONCHIAL:
1. Collect 2 -5 mL of bronchial washings, transbronchial biopsies, or bronchial secretions aspirated through the inner channel of the bronchoscope with or without irrigating solutions.
2. Submit specimen in a sterile, screw-capped, collection container.
NOTE: When using the Sherwood trap collection container, replace the cap attached to the collection tubing with the second screw cap contained in the package to prevent specimen leakage during transport. Discard the primary cap with tubing as biohazard waste.

ENDOTRACHEAL ASPIRATE:
1. Collect specimen (2 - 5 mL) of pulmonary secretions from an endotracheal tube
2. Submit specimen in sterile screw capped collection container.
NOTE: When using the Sherwood trap collection container, replace the cap attached to the collection tubing with the second screw cap contained in the package to prevent specimen leakage during transport. Discard the primary cap with tubing as biohazard waste.

SPUTUM:
1. Instruct patient to brush his/her teeth and/or rinse mouth well with water.
2. Have patient remove dentures.
3. Instruct the patient to take a deep breath, hold it momentarily, and cough deeply and vigorously into the container. Be sure patient understands the difference between sputum and saliva
4. Collect 2 - 5 mL of sputum from an early morning deep cough in a screw-capped, sterile container.
5. For patients with a nonproductive cough, ultrasonic nebulization with 10% saline, hydration, chest physiotherapy, and postural drainage may be ordered.
6. Submit specimen in sterile, screw-capped container.
NOTE: The specimen should be a single, first morning, "deep cough" sputum specimen, and the patient should not have eaten prior to collection.

PLEURAL FLUID:
1. Decontamination skin with povidone iodine.
2. Using sterile technique aspirate pleural fluid with needle and syringe.
3. Send 10-mL of pleural fluid in a screw-capped, sterile tube.

TISSUE:
1. Aseptically obtain 5-10 cubic mm piece of tissue. Avoid collecting tissue contaminated with flora from adjacent mucosal membranes.
2. Do not allow tissue to dry out.
3. Submit in an Anaerobic Transport Tube (AS Tube) or screw-capped, sterile specimen cup (if tissue is too large for AS Tube).
Note: FORMALIN FIXED TISSUE IS NOT ACCEPTABLE.

Blood:
Order the CULTURE BACTERIA BLOOD and indicate LEGIONELLA on the order comments to notify the laboratory that Legionella is suspected.
1. Collect blood cultures prior to initiation of antimicrobial therapy when possible.
2. Throughly cleanse venipuncture site with 2%-4% chlorhexidine gluconate/70% alcohol. Scrub the skin for 15 seconds. Unlike Povidone Iodine (or 2% tincture of iodine), it is not necessary to move in concentric circles to the periphery. Instead, the mechanism of action of chlorhexidine gluconate is based more on friction against the skin.
3. Allow to dry. DO NOT palpate site after preparation.
4. Draw 10 mL of blood for an adult using the Isolator Vacutainer Tube System or 1.5 mL of blood for a pediatric patient using the Pediatric Isolator Vacutainer Tube System. The routine blood culture kit may also be used.

GENERAL NOTES:
1. CULTURE SITE IS REQUIRED ON REQUEST FORM FOR PROCESSING.
2. Isolation of Legionella requires special media and a longer incubation time than most bacterial cultures.

DELIVER IMMEDIATELY TO MICROBIOLOGY IN A TIGHTLY SEALED CONTAINER WITH NO EXTERNAL SPILLAGE.

Causes for Rejction:
1. Specimen container leaking.

Used for diagnosis of infections with Legionella species.

Legionnaires’ Disease, named after the outbreak in 1976 at the American Legion convention in Philadelphia, is caused by Legionella pneumophila and is characterized as an acute febrile respiratory illness ranging in severity from mild illness to fatal pneumonia. The disease occurs in both epidemic and endemic forms and sporadic cases are not easily differentiated from other respiratory infections by clinical symptoms. An estimated 25,000 to 100,000 cases of Legionella infection occur in the United States annually. The resulting mortality rate, ranging from 25% to 40% can be lowered if the disease is diagnosed rapidly and appropriate antimicrobial therapy is instituted early.  

Known risk factors for Legionella infection include immunosuppression, cigarette smoking, alcohol consumption, and concomitant pulmonary disease. The young and the elderly are particularly susceptible.

The most sensitive method to detect Legionella in clinical specimens is by direct culture onto selective and non-selective media.  The sensitivity of culture is approximately 90%.

Legionella are aerobic, fastidious, gram-negative bacilli that require specialized media containing the amino acid L-cysteine for growth.  In clinical specimens where abundant flora are present (e.g., sputum), Legionella can be easily overgrown or inhibited.
There are currently 29 named species of Legionella. These organisms may cause either community-acquired or nosocomially-acquired pneumonia.  Legionella pneumophila is responsible for 80-90% of reported cases of Legionella infection.  Legionella pneumophila is comprised of 14 serogroups with serogroup 1 being responsible for approximately 80-85% of all human cases.  Legionella micdadei (Pittsburg Pneumonia Agent) is the second most commonly isolated species and frequently causes pneumonia in immunocompromised hosts.

Gram stain and aerobic culture selective for recovery of Legionella species. 

Includes identification of Legionella species. Isolates are sent to North Carolina State Laboratory of Public Health for speciation and sero-grouping.

Isolation of Legionella requires special media and a longer incubation time than most bacterial cultures.

Current methods for the laboratory detection of pneumonia caused by Legionella require a respiratory sample, a urine sample antigen detection, or paired sera for an acute diagnosis. One of the presenting sign’s of patients with Legionnaires’ Disease is the relative lack of productive sputum.  In many patients, this necessitates the use of an invasive procedure to obtain a respiratory sample.  

Diagnosis by serological techniques is usually retrospective in nature. Refer to "Legionella Antigen, Urine" for additional information.