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 | |
| Publication status | Published - Jun 1996 |
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Keywords
- DHFR
- DNA sequence
- Pneumocystis carinii
- PVR
ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
Cite this
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
}
TY - JOUR
T1 - Non specific PCR products using rat-derived Pneumocystis carinii dihydrofolate reductase gene-specific primers in DNA amplification of human respiratory samples
AU - Ortona, Elena
AU - Margutti, Paola
AU - De Luca, Andrea
AU - Peters, Sarah E.
AU - Wakefield, Ann E.
AU - Tamburrini, Enrica
AU - Mencarini, Paola
AU - Visconti, Elena
AU - Siracusano, Alessandra
PY - 1996/6
Y1 - 1996/6
N2 - 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.
AB - 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.
KW - DHFR
KW - DNA sequence
KW - Pneumocystis carinii
KW - PVR
UR - http://www.scopus.com/inward/record.url?scp=0029890467&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029890467&partnerID=8YFLogxK
U2 - 10.1006/mcpr.1996.0026
DO - 10.1006/mcpr.1996.0026
M3 - Article
C2 - 8799372
AN - SCOPUS:0029890467
VL - 10
SP - 187
EP - 190
JO - Molecular and Cellular Probes
JF - Molecular and Cellular Probes
SN - 0890-8508
IS - 3
ER -