Microbiological diagnosis of empyema in children: comparative evaluations by culture, polymerase chain reaction, and pneumococcal antigen detection in pleural fluids.
BACKGROUND
Pleural empyema is an increasingly reported complication of pneumonia in children. Microbiological diagnostic tests for empyema by culture frequently have false-negative results due to previous administration of antibiotics. Molecular diagnosis by broad-range 16S ribosomal DNA (rDNA) polymerase chain reaction (PCR) and rapid pneumococcal antigen detection are reliable tools, but their diagnostic value has not been clearly established for pleural fluid samples. Pneumococcal antigen detection has only been validated for urine and cerebrospinal fluid samples.


METHODS
Over 4 years, pleural fluid specimens were collected from 78 children with pleural empyema. Standard culture, pneumococcal antigen detection by latex agglutination (Pastorex; Bio-Rad) and immunochromatographic testing (Binax NOW Streptococcus pneumoniae), and 16S rDNA PCR were performed on these specimens. Pneumococcal identification by 16S rDNA PCR and sequencing was confirmed by pneumolysin PCR.


RESULTS
Of the 78 cases of pleural empyema, 60 (77%) were microbiologically documented by culture or 16S rDNA PCR. Of the 40 pneumococcal empyema cases, 17 (43%) were only diagnosed by PCR and 23 with PCR and culture. The sensitivity and specificity of the latex antigen detection (with the use of culture and/or PCR as the test standard) were 90% and 95%, respectively. The immunochromatographic test detected pneumococcal antigens in 3 additional specimens for which latex agglutination results were negative, thereby increasing the sensitivity of antigen detection.


CONCLUSIONS
Pneumococcal antigen detection in pleural fluid specimens from children provides a rapid and sensitive method of diagnosis of pneumococcal empyema, which can be confirmed by specific pneumolysin PCR when culture results are negative. Broad-range 16S rDNA PCR has value in detecting bacterial agents responsible for culture-negative pleural empyema.
Le Monnier
Alban
A
Assistance Publique-Hopitaux de Paris, Paris, France.
Carbonnelle
Etienne
E
Zahar
Jean-Ralph
JR
Le Bourgeois
Muriel
M
Abachin
Eric
E
Quesne
Gilles
G
Varon
Emmanuelle
E
Descamps
Philippe
P
De Blic
Jacques
J
Scheinmann
Pierre
P
Berche
Patrick
P
Ferroni
Agnes
A
D000942:Antigens, Bacterial / Q000032:analysis* / Q000235:genetics; D001483:Base Sequence; D002648:Child; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics; D004282:Documentation; D004653:Empyema / Q000175:diagnosis*; D016724:Empyema, Pleural / Q000175:diagnosis*; D006801:Humans; D010297:Paris; D010996:Pleural Effusion / Q000382:microbiology*; D011008:Pneumococcal Infections / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results
D000942:Antigens, Bacterial; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S