Abstract
Streptococcus pneumoniae is the most important cause of childhood pneumonia and empyema, yet the diagnosis of pneumococcal infections by conventional methods is challenging. In this study, the clinical value of the pneumolysin-targeted real-time polymerase chain reaction (PCR) method for the diagnosis of pneumococcal pneumonia and empyema was evaluated with 33 whole blood samples and 12 pleural fluid samples. The analytical sensitivity of the PCR assay was 4 fg of pneumococcal DNA, corresponding to two genome equivalents of pneumococcal DNA per reaction. The PCR assay correctly detected all clinical isolates of S. pneumoniae tested, whereas all nonpneumococcal bacterial organisms tested were negative by PCR. In a clinical trial, S. pneumoniae was detected by PCR in the pleural fluid of 75% of children with empyema, increasing the detection rate of pneumococcus almost tenfold that of pleural fluid culture. However, in whole blood samples, PCR detected S. pneumoniae in only one child with pneumonia and one child with pneumococcal empyema and failed to detect S. pneumoniae in three children with blood cultures positive for S. pneumoniae. The present data indicate that pneumolysin-targeted real-time PCR of pleural fluid is a valuable method for the etiologic diagnosis of pneumococcal empyema in children. The ease and rapidity of the LightCycler technology (Roche Diagnostics, Mannheim, Germany) make real-time PCR an applicable tool for routine diagnostics. In the evaluation of blood samples, blood culture remains the superior method for the diagnosis of bacteremic pneumococcal disease.
Similar content being viewed by others
References
Wubbel L, Muniz L, Ahmed A, Trujillo M, Carubelli C, McCoig C, Abramo T, Leinonen M, McCracken GH (1999) Etiology and treatment of community-acquired pneumonia in ambulatory children. Pediatr Infect Dis J 18:98–104
Juven T, Mertsola J, Waris M, Leinonen M, Meurman O, Roivainen R, Eskola J, Saikku P, Ruuskanen O (2000) Etiology of community-acquired pneumonia in 254 hospitalized children. Pediatr Infect Dis J 19:293–298
Vuori-Holopainen E, Salo E, Saxen H, Hedman K, Hyypia T, Lahdenpera R, Leinonen M, Tarkka E, Vaara M, Peltola H (2002) Etiological diagnosis of childhood pneumonia by use of transthoracic needle aspiration and modern microbiological methods. Clin Infect Dis 34:583–590
Michelow IC, Olsen K, Lozano J, Rollins NK, Duffy LB, Ziegler T, Kauppila J, Leinonen M, McCracken GH (2004) Epidemiology and clinical characteristics of community-acquired pneumonia in hospitalized children. Pediatrics 113:701–707
Byington CL, Spencer LY, Johnson TA, Pavia AT, Allen D, Mason EO, Kaplan S, Carroll KC, Daly JA, Christenson JC, Samore MH (2002) An epidemiological investigation of a sustained high rate of pediatric parapneumonic empyema: risk factors and microbiological associations. Clin Infect Dis 34:434–440
Eastham KM, Freeman R, Kearns AM, Eltringham G, Clark J, Leeming J, Spencer DA (2004) Clinical features, aetiology and outcome of empyema in children in the north east of England. Thorax 59:522–525
Gendrel D, Raymond J, Moulin F, Iniguez JL, Ravilly S, Habib F, Lebon P, Kalifa G (1997) Etiology and response to antibiotic therapy of community-acquired pneumonia in French children. Eur J Clin Microbiol Infect Dis 16:388–391
Thomson AH, Hull J, Kumar MR, Wallis C, Balfour Lynn IM, on behalf of the British Paediatric Respiratory Society Empyema Study Group (2002) Randomised trial of intrapleural urokinase in the treatment of childhood empyema. Thorax 57:343–347
Dominguez J, Blanco S, Rodrigo C, Azuara M, Gali N, Mainou A, Esteve A, Castellvi A, Prat C, Matas L, Ausina V (2003) Usefulness of urinary antigen detection by an immunochromatographic test for diagnosis of pneumococcal pneumonia in children. J Clin Microbiol 41:2161–2163
Neuman MI, Harper MB (2003) Evaluation of a rapid urine antigen assay for the detection of invasive pneumococcal disease in children. Pediatrics 112:1279–1282
Korppi M, Koskela M, Jalonen E, Leinonen M (1992) Serologically indicated pneumococcal respiratory infection in children. Scand J Infect Dis 24:437–443
Korppi M, Leinonen M (1998) Pneumococcal immune complexes in the diagnosis of lower respiratory infections in children. Pediatr Infect Dis J 17:992–995
Michelow IC, Lozano J, Olsen K, Goto C, Rollins NK, Ghaffar F, Rodriguez-Cerrato V, Leinonen M, McCracken GH (2002) Diagnosis of Streptococcus pneumoniae lower respiratory infection in hospitalized children by culture, polymerase chain reaction, serological testing, and urinary antigen detection. Clin Infect Dis 34:E1–E11
Saukkoriipi A, Palmu A, Kilpi T, Leinonen M (2002) Real-time quantitative PCR for the detection of Streptococcus pneumoniae in the middle ear fluid of children with acute otitis media. Mol Cell Probes 16:385–390
Rantala M, Huikko S, Huovinen P, Jalava J, and the Finnish Study Group for Antimicrobial Resistance (2005) Prevalence and molecular genetics of macrolide resistance among Streptococcus pneumoniae isolates collected in Finland in 2002. Antimicrob Agents Chemother 49:4180–4184
Kwok S, Higuchi R (1989) Avoiding false positives with PCR. Nature 339:237–238
Saglani S, Harris KA, Wallis C, Hartley JC (2005) Empyema: the use of broad range 16S rDNA PCR for pathogen detection. Arch Dis Child 90:70–73
Farrell DJ, Morrissey I, Bakker S, Felmingham D (2001) Detection of macrolide resistance mechanisms in Streptococcus pneumoniae and Streptococcus pyogenes using a multiplex rapid cycle PCR with microwell-format probe hybridization. J Antimicrob Chemother 48:541–544
Nagai K, Shibasaki Y, Hasegawa K, Davies TA, Jakobs MR, Ubukata K, Appelbaum PC (2001) Evaluation of PCR primers to screen for Streptococcus pneumoniae isolates and beta-lactam resistance, and to detect common macrolide resistance determinants. J Antimicrob Chemother 48:915–918
Kearns AM, Graham C, Burdess D, Heatherington J, Freeman R (2002) Rapid real-time PCR for determination of penicillin susceptibility in pneumococcal meningitis, including culture-negative cases. J Clin Microbiol 40:682–684
Lonks JR, Garau J, Gomez L, Xercavins M, Ochoa de Echagüen A, Gareen IF, Reiss PT, Medeiros AA (2002) Failure of macrolide antibiotic treatment in patients with bacteremia due to erythromycin-resistant Streptococcus pneumoniae. Clin Infect Dis 35:556–564
Jacobs MR, Johnson CE (2003) Macrolide resistance: an increasing concern for treatment failure in children. Pediatr Infect Dis J 22(Suppl 8):131–138
Rudolph KM, Parkinson AJ, Black CM, Mayer LW (1993) Evaluation of polymerase chain reaction for diagnosis of pneumococcal pneumonia. J Clin Microbiol 31:2661–2666
Salo P, Ortqvist A, Leinonen M (1995) Diagnosis of bacteremic pneumococcal pneumonia by amplification of pneumolysin gene fragment in serum. J Infect Dis 171:479–482
Dagan R, Shriker O, Hazan I, Leibovitz E, Greenberg D, Schlaeffer F, Levy R (1998) Prospective study to determine clinical relevance of detection of pneumococcal DNA in sera of children by PCR. J Clin Microbiol 36:669–673
Toikka P, Nikkari S, Ruuskanen O, Leinonen M, Mertsola J (1999) Pneumolysin PCR-based diagnosis of invasive pneumococcal infection in children. J Clin Microbiol 37:633–637
Dominguez J, Gali N, Matas L, Pedroso P, Blanco S, Gimenez M, Prat C, Sopena N, Sabria M, Ausina V (2001) PCR detection of Streptococcus pneumoniae DNA in serum samples for pneumococcal pneumonia diagnosis. Clin Microbiol Infect 7:164–166
Murdoch DR, Anderson TP, Beynon KA, Chua A, Fleming AM, Laing RTR, Town GI, Mills GD, Chambers ST, Jennigs LC (2003) Evaluation of a PCR assay for detection of Streptococcus pneumoniae in respiratory and nonrespiratory samples from adults with community-acquired pneumonia. J Clin Microbiol 41:63–66
van Haeften R, Palladino S, Kay I, Keil T, Heath C, Waterer GW (2003) A quantitative LightCycler PCR to detect Streptococcus pneumoniae in blood and CSF. Diagn Microbiol Infect Dis 47:407–414
Sullivan TD, LaScolea LJ Jr, Neter E (1982) Relationship between the magnitude of bacteremia in children and the clinical disease. Pediatrics 69:699–702
Bell LM, Alpert G, Campos JM, Plotkin SA (1985) Routine quantitative blood cultures in children with Haemophilus influenzae or Streptococcus pneumoniae bacteremia. Pediatrics 76:901–904
Toikka P, Virkki R, Mertsola J, Ashorn P, Eskola J, Ruuskanen O (1999) Bacteremic pneumococcal pneumonia in children. Clin Infect Dis 29:568–572
Acknowledgements
We thank Tiina Haarala and Tuula Randell for skilful technical assistance.
This study was financially supported by the Finnish Cultural Foundation, the Finnish Foundation for Pediatric Research, the Maud Kuistila Foundation, and the Turku University Foundation.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lahti, E., Mertsola, J., Kontiokari, T. et al. Pneumolysin polymerase chain reaction for diagnosis of pneumococcal pneumonia and empyema in children. Eur J Clin Microbiol Infect Dis 25, 783–789 (2006). https://doi.org/10.1007/s10096-006-0225-9
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10096-006-0225-9