Detection of Pneumocystis carinii in tracheal aspirates of intubated patients using calcofluor-white (Fungi-Fluor) and immunofluorescence antibody (Genetic Systems) stains

Crit Care Med. 1997 Jun;25(6):948-52. doi: 10.1097/00003246-199706000-00009.

Abstract

Objective: To compare the detection rate of Pneumocystis carinii in endotracheal aspirates with that rate in bronchoalveolar lavage fluid, using calcofluor-white (Fungi-Fluor) and immunofluorescence antibody (Genetic Systems) staining methods.

Design: Prospective, consecutive cases.

Setting: Medical intensive care unit at Ben Taub General Hospital.

Patients: Thirty-one intubated patients admitted with respiratory failure and suspected P. carinii pneumonia.

Interventions: An endotracheal aspirate specimen was obtained after maximally advancing a closed-system suction catheter, instilling aliquot portions of saline, and suctioning the lavage fluid. This procedure was followed within 30 mins by fiberoptic bronchoscopy and bronchoalveolar lavage.

Measurements and main results: Endotracheal aspirate and bronchoalveolar lavage specimens from each patient were mixed with Saccomano's fixative, blended, and centrifuged. Using a modified method for P. carinii cysts, the sediment was stained with the test calcofluor-white stain Solution A and the test antibody stain. The test antibody stain on the bronchoalveolar lavage specimens was positive for P. carinii for 13 patients and was used as the standard for comparison. In the endotracheal aspirate specimens, the test antibody stain detected 12 (92%) P. carinii-positive patients while the test calcofluor-white stain detected ten (77%) P. carinii-positive patients.

Conclusions: We described a simple method for obtaining, processing, and staining endotracheal aspirate specimens for P. carinii. Obtaining an endotracheal aspirate specimen did not require specially trained personnel or a specialized and more expensive catheter, and was not associated with any complications.

Publication types

  • Comparative Study

MeSH terms

  • Benzenesulfonates*
  • Contrast Media*
  • Fluorescent Antibody Technique, Direct*
  • Fluorescent Dyes*
  • Humans
  • Intubation, Intratracheal
  • Pneumocystis / isolation & purification*
  • Pneumonia, Pneumocystis / microbiology
  • Prospective Studies
  • Respiratory Insufficiency / microbiology
  • Suction
  • Trachea / microbiology

Substances

  • Benzenesulfonates
  • Contrast Media
  • Fluorescent Dyes
  • C.I. Fluorescent Brightening Agent 28