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J Clin Microbiol. Nov 1994; 32(11): 2667–2670.
PMCID: PMC264139

Comparison of PCR assay with bacterial culture for detecting Streptococcus pneumoniae in middle ear fluid of children with acute otitis media.


We have studied etiological diagnosis of acute otitis media (AOM) by comparing a newly developed pneumococcal PCR for Streptococcus pneumoniae to bacterial culture with 180 middle ear fluid (MEF) samples of 125 children with 125 episodes of AOM. For pneumococcal PCR assay, DNA from MEF samples was extracted by phenol-chloroform. The outer primers used amplified a 348-bp region of the pneumolysin gene, and the inner primers amplified a 208-bp region. S. pneumoniae was cultured in 33 (18%) samples, and pneumolysin PCR was positive for 51 (28%) of 180 MEF samples. Only 2 of 21 PCR-positive, S. pneumoniae culture-negative samples were positive for other otitis pathogens. By combining MEF culture and PCR results, 54 (30%) of 180 MEF samples had evidence of pneumococcal etiology. In conclusion, pneumolysin PCR is a sensitive and specific new method to study pneumococcal involvement in MEF samples of children with AOM.

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  • Donofrio JC, Coonrod JD, Davidson JN, Betts RF. Detection of influenza A and B in respiratory secretions with the polymerase chain reaction. PCR Methods Appl. 1992 May;1(4):263–268. [PubMed]
  • He Q, Mertsola J, Soini H, Skurnik M, Ruuskanen O, Viljanen MK. Comparison of polymerase chain reaction with culture and enzyme immunoassay for diagnosis of pertussis. J Clin Microbiol. 1993 Mar;31(3):642–645. [PMC free article] [PubMed]
  • Karma P, Palva T, Kouvalainen K, Kärjä J, Mäkelä PH, Prinssi VP, Ruuskanen O, Launiala K. Finnish approach to the treatment of acute otitis media. Report of the Finnish Consensus Conference. Ann Otol Rhinol Laryngol Suppl. 1987 Mar-Apr;129:1–19. [PubMed]
  • Khan G, Kangro HO, Coates PJ, Heath RB. Inhibitory effects of urine on the polymerase chain reaction for cytomegalovirus DNA. J Clin Pathol. 1991 May;44(5):360–365. [PMC free article] [PubMed]
  • Kwok S, Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. [PubMed]
  • Leinonen MK. Detection of Pneumococcal Capsular polysaccharide antigens by latex agglutination, counterimmunoelectrophoresis, and radioimmunoassay in middle ear exudates in acute otitis media. J Clin Microbiol. 1980 Feb;11(2):135–140. [PMC free article] [PubMed]
  • Luotonen J. Streptococcus pneumoniae and Haemophilus influenzae in nasal cultures during acute otitis media. Acta Otolaryngol. 1982 Mar-Apr;93(3-4):295–299. [PubMed]
  • Luotonen J, Herva E, Karma P, Timonen M, Leinonen M, Mäkelä PH. The bacteriology of acute otitis media in children with special reference to Streptococcus pneumoniae as studied by bacteriological and antigen detection methods. Scand J Infect Dis. 1981;13(3):177–183. [PubMed]
  • Nolte FS, Metchock B, McGowan JE, Jr, Edwards A, Okwumabua O, Thurmond C, Mitchell PS, Plikaytis B, Shinnick T. Direct detection of Mycobacterium tuberculosis in sputum by polymerase chain reaction and DNA hybridization. J Clin Microbiol. 1993 Jul;31(7):1777–1782. [PMC free article] [PubMed]
  • Okamoto Y, Kudo K, Ishikawa K, Ito E, Togawa K, Saito I, Moro I, Patel JA, Ogra PL. Presence of respiratory syncytial virus genomic sequences in middle ear fluid and its relationship to expression of cytokines and cell adhesion molecules. J Infect Dis. 1993 Nov;168(5):1277–1281. [PubMed]
  • Paton AW, Paton JC, Lawrence AJ, Goldwater PN, Harris RJ. Rapid detection of respiratory syncytial virus in nasopharyngeal aspirates by reverse transcription and polymerase chain reaction amplification. J Clin Microbiol. 1992 Apr;30(4):901–904. [PMC free article] [PubMed]
  • Paton JC, Lock RA, Hansman DJ. Effect of immunization with pneumolysin on survival time of mice challenged with Streptococcus pneumoniae. Infect Immun. 1983 May;40(2):548–552. [PMC free article] [PubMed]
  • Qvarnberg Y, Holopainen E, Palva T. Aspiration cytology in acute otitis media. Acta Otolaryngol. 1984 May-Jun;97(5-6):443–449. [PubMed]
  • Rudolph KM, Parkinson AJ, Black CM, Mayer LW. Evaluation of polymerase chain reaction for diagnosis of pneumococcal pneumonia. J Clin Microbiol. 1993 Oct;31(10):2661–2666. [PMC free article] [PubMed]
  • Soini H, Skurnik M, Liippo K, Tala E, Viljanen MK. Detection and identification of mycobacteria by amplification of a segment of the gene coding for the 32-kilodalton protein. J Clin Microbiol. 1992 Aug;30(8):2025–2028. [PMC free article] [PubMed]
  • Walker JA, Allen RL, Falmagne P, Johnson MK, Boulnois GJ. Molecular cloning, characterization, and complete nucleotide sequence of the gene for pneumolysin, the sulfhydryl-activated toxin of Streptococcus pneumoniae. Infect Immun. 1987 May;55(5):1184–1189. [PMC free article] [PubMed]

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