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

E Ortona; P Margutti; A De Luca; S E Peters; A E Wakefield; E Tamburrini; P Mencarini; E Visconti; A Siracusano

doi:10.1006/mcpr.1996.0026

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

Mol Cell Probes. 1996 Jun;10(3):187-90. doi: 10.1006/mcpr.1996.0026.

Authors

E Ortona¹, P Margutti, A De Luca, S E Peters, A E Wakefield, E Tamburrini, P Mencarini, E Visconti, A Siracusano

Affiliation

¹ Laboratory of Immunology, Istituto Superiore di Sanità, Rome, Italy.

PMID: 8799372
DOI: 10.1006/mcpr.1996.0026

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 non-specific 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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
Bronchoalveolar Lavage Fluid / chemistry
DNA Primers
Humans
Molecular Sequence Data
Pneumocystis / enzymology*
Polymerase Chain Reaction / methods*
Tetrahydrofolate Dehydrogenase / genetics*

Substances

DNA Primers
Tetrahydrofolate Dehydrogenase