Non specific PCR products using rat-derived Pneumocystis carinii dihydrofolate reductase gene-specific primers in DNA amplification of human respiratory samples

Elena Ortona, Paola Margutti, Andrea De Luca, Sarah E. Peters, Ann E. Wakefield, Enrica Tamburrini, Paola Mencarini, Elena Visconti, Alessandra Siracusano

Research output: Contribution to journalArticlepeer-review

Abstract

DNA amplification using dihydrofolate reductase (DHFR) primers in bronchoalveolar lavage fluids (BALFs) from patients with Pneumocystis carinii (PC) pneumonia yielded low sensitivity and specificity. Amplified products of BALFs from an AIDS patient without PC pneumonia and five patients with PC pneumonia were cloned and sequenced. All samples showed the same sequence without any homology with DHFR cDNA of rat PC, or with any DHFR sequences in databases at the DNA or amino acid level. The data demonstrate that these DHFR primers amplify a nonspecific region of DNA with a sequence unrelated to the human PC DHFR gene both in PC positive and in PC negative samples. This finding precludes the use of these DHFR primers for the diagnosis of PC pneumonia in respiratory specimens.

Original languageEnglish
Pages (from-to)187-190
Number of pages4
JournalMolecular and Cellular Probes
Volume10
Issue number3
DOIs
Publication statusPublished - Jun 1996

Keywords

  • DHFR
  • DNA sequence
  • Pneumocystis carinii
  • PVR

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

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