title: Non specific PCR products using rat-derived Pneumocystis carinii dihydrofolate reductase gene-specific primers in DNA amplification of human respiratory samples.
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.
authors.affiliation: Laboratory of Immunology, Istituto Superiore di Sanità, Rome, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Ortona
authors.forename: P
authors.initials: P
authors.lastname: Margutti
authors.forename: A
authors.initials: A
authors.lastname: De Luca
authors.forename: S E
authors.initials: SE
authors.lastname: Peters
authors.forename: A E
authors.initials: AE
authors.lastname: Wakefield
authors.forename: E
authors.initials: E
authors.lastname: Tamburrini
authors.forename: P
authors.initials: P
authors.lastname: Mencarini
authors.forename: E
authors.initials: E
authors.lastname: Visconti
authors.forename: A
authors.initials: A
authors.lastname: Siracusano
mesh_terms: D001483:Base Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D017931:DNA Primers; D006801:Humans; D008969:Molecular Sequence Data; D011010:Pneumocystis / Q000201:enzymology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*
chemical_list: D017931:DNA Primers; D013762:Tetrahydrofolate Dehydrogenase