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).