Tritiated Thymidine BeLPT: Bronchoalveolar Lavage (BAL) Specimen

DOE-SPEC-1142-2001

7. TRITIATED THYMIDINE BeLPT: BRONCHOALVEOLAR LAVAGE (BAL)

SPECIMEN

7.1.

SAMPLE COLLECTION. For the BAL-LPT, the specimen is a saline lavage

obtained during bronchoscopy. It is recommended that samples be taken from the

wedged position in lingula or right middle lobe. The lavage is preformed before the

biopsy to avoid contamination with blood. A minimum of 480 ml normal saline is

instilled in two 240 ml aliquots. Gentle aspiration occurs after each aliquot is

instilled. Fluid recovery usually varies from 50-70%. The specimen must be

transported to the laboratory immediately. Temperature should be kept between 15 to

25C using insulation if necessary.

Specimens that cannot be set up for proliferation testing that day must be suspended

in RPMI-1640 with antibiotics and processed within 36 hours (no more than one

night should pass before the cells are placed into culture for proliferation assays). The

test requires a minimum of 2.5 x 106 cells and 20 x 106 are recommended.

7.2.

SAMPLE PREPARATION. BAL preparations from normal individuals usually

consist of over 90% alveolar macrophages. However, in beryllium disease higher

percentages of lymphocytes are typically observed. BAL fluid is centrifuged, the cells

re-suspended in culture medium, counted, and exposed to beryllium for 3 and 5 days.

The presence of immuno-competent T lymphocytes in the preparation is usually

ascertained by testing with Candida albicans, a common recall antigen, although

comparable universal antigens/mitogens may be employed to the same end. Tritiated

thymidine is included for the last 6-18 hrs. of culture and the assay is terminated by

harvesting cells onto filter paper and determining the radioactive content in a

scintillation or gas ionization counter.

7.3.

SAMPLE FORM

7.3.1. PATIENT NAME: BIRTH DATE:

7.3.2. HOSPITAL#

7.3.3. DATE AND TIME OF TEST: SW:

7.3.4. SPECIMEN #

7.3.5. LAVAGE VOLUME- ml IN:

7.3.6. ml OUT: %Return:

7.3.7. CELL YIELD: Total number of cells recovered/ml lavage fluid out