Pneumocystis carinii in bronchoalveolar lavage and induced sputum: detection with a nested polymerase chain reaction.
To evaluate polymerase chain reaction (PCR) for detection of Pneumocystis carinii, 117 bronchoalveolar lavage (BAL) specimens, from HIV-infected patients undergoing a diagnostic bronchoscopy, were processed and a nested PCR, followed by Southern blot and hybridization with a P32-labelled probe was performed. The sensitivity and specificity were 85 and 100% 934/40 and 77/77) respectively. A non-radioactive labelling system BluGENE was evaluated on all specimens, and found to be as effective as P32-labelling. To increase the speed and convenience of detection, a dot blot system was tested, but sensitivity dropped markedly with this system. A further 33 patients had both induced sputum and bronchoalveolar lavage performed and the induced sputum was analysed using PCR and routine microbiological methods. The PCR sensitivity on induced sputum was equal to that of routine methods. At present the evaluated PCR cannot replace routine microbiological methods for detection of Pneumocystis carinii, on either BAL fluid or induced sputum.
Skøt
J
J
Department of Infectious Diseases, University of Copenhagen, Hvidovre Hospital, Denmark.
Lerche
A G
AG
Kolmos
H J
HJ
Nielsen
J O
JO
Mathiesen
L R
LR
Lundgren
J D
JD
D017088:AIDS-Related Opportunistic Infections / Q000175:diagnosis*; D001483:Base Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D001999:Bronchoscopy; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis; D005188:False Negative Reactions; D006801:Humans; D008969:Molecular Sequence Data; D011010:Pneumocystis / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology*
D017931:DNA Primers; D004271:DNA, Fungal