Development and evaluation of a molecular viability assay for Pneumocystis carinii

Nancy Maher, Sten Vermund, Mark Lasbury, Chao-Hung Lee, Marilyn Bartlett, Thomas R. Unnasch

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Despite recent declines in incidence, Pneumocystis carinii pneumonia (PCP) remains the most commonly occurring opportunistic illness among persons with AIDS in the United States. While P. carinii DNA has been detected in patient respiratory specimens and in air samples collected from various indoor environments housing PCP patients, the viability of these organisms is unknown. For this reason, we have developed and evaluated a molecular viability assay for P. carinii. This method is based upon the detection of P. carinii mRNA by a reverse transcription-PCR that employs specific primers from a member of the heat shock protein 70 family. Under optimal assay conditions, these primers were capable of detecting as few as 100 viable trophozoites as determined by ethidium bromide staining, while no signal was obtained from 106 trophozoites killed by heat, desiccation, or UV radiation. This assay was also capable of distinguishing P. carinii from other common fungi present in the air. Therefore, this molecular viability assay may be useful in conjunction with standard bioaerosol collection devices and procedures for the detection of viable P. carinii collected from various indoor environments. It may also be useful in confirming the presence of viable trophozoites in respiratory specimens collected by noninvasive techniques from putatively infected individuals.

Original languageEnglish
Pages (from-to)1947-1952
Number of pages6
JournalJournal of Clinical Microbiology
Volume38
Issue number5
StatePublished - 2000

Fingerprint

Pneumocystis carinii
Trophozoites
Pneumocystis Pneumonia
Air
Desiccation
HSP70 Heat-Shock Proteins
Ethidium
Reverse Transcription
Acquired Immunodeficiency Syndrome
Fungi
Hot Temperature
Radiation
Staining and Labeling
Equipment and Supplies
Polymerase Chain Reaction
Messenger RNA
DNA
Incidence

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Maher, N., Vermund, S., Lasbury, M., Lee, C-H., Bartlett, M., & Unnasch, T. R. (2000). Development and evaluation of a molecular viability assay for Pneumocystis carinii. Journal of Clinical Microbiology, 38(5), 1947-1952.

Development and evaluation of a molecular viability assay for Pneumocystis carinii. / Maher, Nancy; Vermund, Sten; Lasbury, Mark; Lee, Chao-Hung; Bartlett, Marilyn; Unnasch, Thomas R.

In: Journal of Clinical Microbiology, Vol. 38, No. 5, 2000, p. 1947-1952.

Research output: Contribution to journalArticle

Maher, N, Vermund, S, Lasbury, M, Lee, C-H, Bartlett, M & Unnasch, TR 2000, 'Development and evaluation of a molecular viability assay for Pneumocystis carinii', Journal of Clinical Microbiology, vol. 38, no. 5, pp. 1947-1952.
Maher, Nancy ; Vermund, Sten ; Lasbury, Mark ; Lee, Chao-Hung ; Bartlett, Marilyn ; Unnasch, Thomas R. / Development and evaluation of a molecular viability assay for Pneumocystis carinii. In: Journal of Clinical Microbiology. 2000 ; Vol. 38, No. 5. pp. 1947-1952.
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