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Laboratory implementation of a rapid three-stain technique for detection of microorganisms from lower respiratory specimens.
J Clin Lab Anal. 1996; 10(2):104-9.JC

Abstract

A rapid, cost-effective method for the evaluation of lower respiratory specimen has become increasingly important in the diagnosis of pulmonary diseases in immunocompromised patients. In the past, the technically demanding, time-consuming, and expensive Gomori-methenamine-silver (GMS) stain was the principal means for the evaluation of these specimens. In this study, we compared the GMS stain with a new rapid, three-stain protocol for the evaluation of lower respiratory specimens. Lower respiratory specimens were obtained by bronchoalveolar lavage (BAL). Conventional Wright/Giemsa and Gram stains were utilized, as well as a contemporary strain, calcofluor white (CW). A cell count was performed on the BAL specimens, and cytospins were stained by the three stains. The calcofluor white-stained slides were examined with an epi-fluorescent microscope, whereas the other stains were evaluated with a conventional light microscope. Gomorimethenamine-silver (GMS), acid-fast bacillus (AFB), and Papanicolaou (PAP) stains were performed as controls. Thirty-two BAL procedures were performed in 20 (63%) male patients and 12 (37%) female patients. The clinical diagnosis was pneumonia in 31% of the patients, malignant hematologic disease in 28%, acute respiratory distress syndrome (ARDS) in 9%, and acquired immunodeficiency syndrome (AIDS) in 28%. Of these specimens, 78% were adequate for interpretation and 22% were inadequate. Bacteria were found in 50% (16/32) of all BALs, fungi were found in 9% (3/32), and Pneumocystis carinii was found in 9% (3/32). Gram-positive bacteria were most frequently found in patients with pneumonia (80%, 4/5), whereas P. carinii was identified in patients with AIDS. There were no false-positive results. One CW stain was equivocal for P. carinii due to high fluorescent background. Laboratory implementation of the rapid, three-staining technique was accomplished without difficulty in microbiology and hematology laboratory sections. Specimen evaluation with the rapid staining protocol was technically easy to perform; however, experience in ultraviolet fluorescent microscopy was crucial for interpretation of CW stain. All results were available in 2 hr, cost was reduced by 30%, and the assays were available 7 days/week. Further studies are ongoing to substantiate the sensitivity, specificity, and predictive value of this technique, as well as clinical guidelines for its optimal utilization.

Authors+Show Affiliations

Department of Pathology, Robert C. Byrd Health Sciences Center, West Virginia University, Morgantown 26506, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

8852363

Citation

Maymind, M, et al. "Laboratory Implementation of a Rapid Three-stain Technique for Detection of Microorganisms From Lower Respiratory Specimens." Journal of Clinical Laboratory Analysis, vol. 10, no. 2, 1996, pp. 104-9.
Maymind M, Thomas JG, Abrons HL, et al. Laboratory implementation of a rapid three-stain technique for detection of microorganisms from lower respiratory specimens. J Clin Lab Anal. 1996;10(2):104-9.
Maymind, M., Thomas, J. G., Abrons, H. L., & Riley, R. S. (1996). Laboratory implementation of a rapid three-stain technique for detection of microorganisms from lower respiratory specimens. Journal of Clinical Laboratory Analysis, 10(2), 104-9.
Maymind M, et al. Laboratory Implementation of a Rapid Three-stain Technique for Detection of Microorganisms From Lower Respiratory Specimens. J Clin Lab Anal. 1996;10(2):104-9. PubMed PMID: 8852363.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Laboratory implementation of a rapid three-stain technique for detection of microorganisms from lower respiratory specimens. AU - Maymind,M, AU - Thomas,J G, AU - Abrons,H L, AU - Riley,R S, PY - 1996/1/1/pubmed PY - 2000/6/20/medline PY - 1996/1/1/entrez SP - 104 EP - 9 JF - Journal of clinical laboratory analysis JO - J Clin Lab Anal VL - 10 IS - 2 N2 - A rapid, cost-effective method for the evaluation of lower respiratory specimen has become increasingly important in the diagnosis of pulmonary diseases in immunocompromised patients. In the past, the technically demanding, time-consuming, and expensive Gomori-methenamine-silver (GMS) stain was the principal means for the evaluation of these specimens. In this study, we compared the GMS stain with a new rapid, three-stain protocol for the evaluation of lower respiratory specimens. Lower respiratory specimens were obtained by bronchoalveolar lavage (BAL). Conventional Wright/Giemsa and Gram stains were utilized, as well as a contemporary strain, calcofluor white (CW). A cell count was performed on the BAL specimens, and cytospins were stained by the three stains. The calcofluor white-stained slides were examined with an epi-fluorescent microscope, whereas the other stains were evaluated with a conventional light microscope. Gomorimethenamine-silver (GMS), acid-fast bacillus (AFB), and Papanicolaou (PAP) stains were performed as controls. Thirty-two BAL procedures were performed in 20 (63%) male patients and 12 (37%) female patients. The clinical diagnosis was pneumonia in 31% of the patients, malignant hematologic disease in 28%, acute respiratory distress syndrome (ARDS) in 9%, and acquired immunodeficiency syndrome (AIDS) in 28%. Of these specimens, 78% were adequate for interpretation and 22% were inadequate. Bacteria were found in 50% (16/32) of all BALs, fungi were found in 9% (3/32), and Pneumocystis carinii was found in 9% (3/32). Gram-positive bacteria were most frequently found in patients with pneumonia (80%, 4/5), whereas P. carinii was identified in patients with AIDS. There were no false-positive results. One CW stain was equivocal for P. carinii due to high fluorescent background. Laboratory implementation of the rapid, three-staining technique was accomplished without difficulty in microbiology and hematology laboratory sections. Specimen evaluation with the rapid staining protocol was technically easy to perform; however, experience in ultraviolet fluorescent microscopy was crucial for interpretation of CW stain. All results were available in 2 hr, cost was reduced by 30%, and the assays were available 7 days/week. Further studies are ongoing to substantiate the sensitivity, specificity, and predictive value of this technique, as well as clinical guidelines for its optimal utilization. SN - 0887-8013 UR - https://www.unboundmedicine.com/medline/citation/8852363/Laboratory_implementation_of_a_rapid_three_stain_technique_for_detection_of_microorganisms_from_lower_respiratory_specimens_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0887-8013&date=1996&volume=10&issue=2&spage=104 DB - PRIME DP - Unbound Medicine ER -