Laboratory diagnosis of Pneumocystis carinii infections by PCR directed to genes encoding for mitochondrial 5S and 28S ribosomal RNA

Gurpreet S. Sandhu, Bruce C. Kline, Mark J. Espy, Leslie Stockman, Thomas F. Smith, Andrew Harold Limper

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

PCR with 5S mitochondrial ribosomal RNA (5S) target is a sensitive and specific assay for the detection of Pneumocystis cariniiin clinical specimens from the respiratory tract. We developed an oligonucleotide probe directed to a 200 bp amplicon generated by fungal-specific universal primers that anneals with sequrences specific for P. carinii in the 28S ribosomal RNA gene (28S). Of 50 archived bronchoalveolar lavage (BAL) specimens, 46 of 50 samples (92% agreement) gave the same result (23 positive, 23 negative) by PCR directed to the 5S and 28S assays. Results of calcofluor white staining of BAL smears on slides indicated agreement with the molecular results in 43 of 46 (93.5%) assays. PCR detection of P. carinii by amplification of 28S ribosomal gene target by fungal-specific primers and an organism-specific probe provides an alternate genomic target for the laboratory diagnosis of this organism. Copyright (C) 1999 Elsevier Science Inc.

Original languageEnglish (US)
Pages (from-to)157-162
Number of pages6
JournalDiagnostic Microbiology and Infectious Disease
Volume33
Issue number3
DOIs
StatePublished - Mar 1999

Fingerprint

28S Ribosomal RNA
5S Ribosomal RNA
Pneumocystis Infections
Mitochondrial Genes
Clinical Laboratory Techniques
Pneumocystis carinii
Bronchoalveolar Lavage
Polymerase Chain Reaction
Fungal Genes
Pneumocystis
Oligonucleotide Probes
rRNA Genes
Respiratory System
Staining and Labeling

ASJC Scopus subject areas

  • Infectious Diseases
  • Immunology and Allergy
  • Virology
  • Parasitology
  • Microbiology
  • Immunology
  • Applied Microbiology and Biotechnology

Cite this

Laboratory diagnosis of Pneumocystis carinii infections by PCR directed to genes encoding for mitochondrial 5S and 28S ribosomal RNA. / Sandhu, Gurpreet S.; Kline, Bruce C.; Espy, Mark J.; Stockman, Leslie; Smith, Thomas F.; Limper, Andrew Harold.

In: Diagnostic Microbiology and Infectious Disease, Vol. 33, No. 3, 03.1999, p. 157-162.

Research output: Contribution to journalArticle

Sandhu, Gurpreet S. ; Kline, Bruce C. ; Espy, Mark J. ; Stockman, Leslie ; Smith, Thomas F. ; Limper, Andrew Harold. / Laboratory diagnosis of Pneumocystis carinii infections by PCR directed to genes encoding for mitochondrial 5S and 28S ribosomal RNA. In: Diagnostic Microbiology and Infectious Disease. 1999 ; Vol. 33, No. 3. pp. 157-162.
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