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

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

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

Istituto Nazionale per le Malattie Infettive Lazzaro Spallanzani

Research output: Contribution to journal › Article › peer-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 language	English
Pages (from-to)	187-190
Number of pages	4
Journal	Molecular and Cellular Probes
Volume	10
Issue number	3
DOIs	https://doi.org/10.1006/mcpr.1996.0026
Publication status	Published - Jun 1996

Keywords

DHFR
DNA sequence
Pneumocystis carinii
PVR

ASJC Scopus subject areas

Cell Biology
Molecular Biology

Access to Document

10.1006/mcpr.1996.0026

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Ortona, E., Margutti, P., De Luca, A., Peters, S. E., Wakefield, A. E., Tamburrini, E., Mencarini, P., Visconti, E., & Siracusano, A. (1996). Non specific PCR products using rat-derived Pneumocystis carinii dihydrofolate reductase gene-specific primers in DNA amplification of human respiratory samples. Molecular and Cellular Probes, 10(3), 187-190. https://doi.org/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. / Ortona, Elena; Margutti, Paola; De Luca, Andrea; Peters, Sarah E.; Wakefield, Ann E.; Tamburrini, Enrica; Mencarini, Paola; Visconti, Elena; Siracusano, Alessandra.

In: Molecular and Cellular Probes, Vol. 10, No. 3, 06.1996, p. 187-190.

Research output: Contribution to journal › Article › peer-review

Ortona, E, Margutti, P, De Luca, A, Peters, SE, Wakefield, AE, Tamburrini, E, Mencarini, P, Visconti, E & Siracusano, A 1996, 'Non specific PCR products using rat-derived Pneumocystis carinii dihydrofolate reductase gene-specific primers in DNA amplification of human respiratory samples', Molecular and Cellular Probes, vol. 10, no. 3, pp. 187-190. https://doi.org/10.1006/mcpr.1996.0026

Ortona, Elena ; Margutti, Paola ; De Luca, Andrea ; Peters, Sarah E. ; Wakefield, Ann E. ; Tamburrini, Enrica ; Mencarini, Paola ; Visconti, Elena ; Siracusano, Alessandra. / Non specific PCR products using rat-derived Pneumocystis carinii dihydrofolate reductase gene-specific primers in DNA amplification of human respiratory samples. In: Molecular and Cellular Probes. 1996 ; Vol. 10, No. 3. pp. 187-190.

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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.",

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