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Document 0079
DOCN M9650079
TI Variable efficiency of three primer pairs for the diagnosis of
Pneumocystis carinii pneumonia by the polymerase chain reaction.
DT 9605
AU De Luca A; Tamburrini E; Ortona E; Mencarini P; Margutti P; Antinori A;
Visconti E; Siracusano A; Institute of Clinical Infectious Diseases,
Catholic University,; Rome, Italy.
SO Mol Cell Probes. 1995 Oct;9(5):333-40. Unique Identifier : AIDSLINE
MED/96123416
AB The efficiency of three different primer pairs, complementary to
different Pneumocystis carinii DNA regions, was compared in the
polymerase chain reaction (PCR) for the diagnosis of Pneumocystis
carinii pneumonia (PCP) on bronchoalveolar fluid (BALF) from patients
with AIDS. PCR coupled with dot-blot hybridization (BLOT) using primers
and probe from the mitochondrial 23SrDNA region showed the highest
sensitivity, with a lower detection limit of 0.5-1 organisms
microliter-1. When testing 47 BALF, PCR plus BLOT of the mitochondrial
23SrDNA region showed also the best diagnostic efficiency (97%
sensitivity, 100% specificity). Sensitivity was significantly higher
than with PCR and BLOT of the 5SrDNA region (81.5% sensitivity; P =
0.025, McNemar test); and of the dehydrofolate reductase (DHFR) gene
region (75.6% sensitivity; P = 0.019). Sensitivity was also
significantly higher than indirect immunofluorescence (75.8%
sensitivity; P = 0.008). Using DHFR primers and probe, specificity was
also reduced. The diagnostic sensitivity in clinical specimens
paralleled the detection limit in the standard dilutions. The use of
repeated DNA sequences of proven specificity as target of PCR
amplification favourably influences sensitivity and specificity. This
comparative study demonstrates that primer selection plays a significant
role in the diagnosis of PCP by PCR.
DE AIDS-Related Opportunistic Infections/*DIAGNOSIS Base Sequence
Comparative Study DNA Primers DNA, Mitochondrial/GENETICS DNA,
Ribosomal/GENETICS Human In Situ Hybridization Molecular Sequence
Data Oligonucleotide Probes Pneumocystis carinii/GENETICS/*ISOLATION &
PURIF Pneumonia, Pneumocystis carinii/*DIAGNOSIS *Polymerase Chain
Reaction Reproducibility of Results RNA, Ribosomal, 23S/GENETICS RNA,
Ribosomal, 5S/GENETICS Support, Non-U.S. Gov't Tetrahydrofolate
Dehydrogenase/GENETICS JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).