PCR-based detection of bacterial DNA after antimicrobial treatment is indicative of persistent, viable bacteria in the chinchilla model of otitis media

Post, J. Christopher; Aul, Jerome J.; White, Gregory J.; Wadowsky, Robert M.; Zavoral, Thomas; Tabari, Reza; Kerber, Brenda; Doyle, William J.; Ehrlich, Garth D.

doi:10.1016/s0196-0709(96)90005-8

Cited by 113 publications

(97 citation statements)

References 8 publications

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“…Trabalhos em animais esclareceram, ao nosso ver, este que é o questionamento principal acerca da aplicabilidade da PCR na investigação da origem infecciosa das doenças da orelha média. Usando o modelo de otite média em chinchilas, Post et al 18 demonstraram que DNA bacteriano purificado e DNA de bactérias inativadas pelo calor não persistiam numa forma amplificável por mais de 3 dias após a inoculação. Além disso, bactérias sensíveis a antibióticos tornaram-se não-cultiváveis após o terceiro dia de tratamento, mas o DNA permaneceu amplificável por 3 semanas.…”

Section: Discussionunclassified

Prevalence of bacteria in children with otitis media with effusion

Pereira¹,

Pereira²,

Cantarelli³

et al. 2004

J Pediatr (Rio J)

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Artigo submetido em 12.05.03, aceito em 15.10.03. AbstractObjectives: 1) To determine the prevalence of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in middle ear effusions of children with otitis media with effusion undergoing myringotomy; 2) to compare the results obtained by culture and PCR; and 3) to determine the susceptibility of bacterial isolates to penicillin. Methods:We analyzed 128 middle ear effusion specimens from 75 children (age = 11 months to 10 years; mean = 34.7 months). Patients with recurrent otitis media had documented middle ear effusion for > 6 weeks, and chronic otitis media with effusion for > 3 months. The patients had no signs of acute otitis media or respiratory tract infection and were not on antibiotic therapy. Aspiration was done through tympanocentesis with an Alden-Senturia trap. Bacteriological studies were initiated less than 15 minutes after specimen collection. Part of the sample was stored at -20 o C for later multiplex PCR analysis. Statistical analysis employed McNemars χ 2 test.Results: Bacteria were cultured in 32 (25.1%) out of 128 samples and the pathogens under investigation were found in 25 (19.6%). PCR was positive for bacteria in 73 (57.0%) specimens: 50 (39.1%) for H. influenzae, 16 (12.5%) for S. pneumoniae, and 13 (10.2%) for M. catarrhalis. All the culture-positive samples were PCR-positive, but 48 (65.7%) of the PCR-positive specimens were culture-negative. PCR was significantly more sensitive than culture (p < 0.01) to identify bacteria. Resistance to penicillin was as follows: M. catarrhalis = 100%; S. pneumoniae = 62.5% and H. influenzae = 23% of the isolates. Conclusions:The prevalence of bacteria in otitis media with effusion in a group of Brazilian children was similar to that reported for other countries. H. influenzae was the most frequent microorganism observed. This suggests that bacteria may play a role in the pathogenesis of otitis media with effusion. In addition, PCR was more sensitive to detect bacteria in middle ear effusion as compared to conventional culture methods. Penicillin resistance was similar to that reported for other countries for pneumococci and moraxella, but beta-lactamase production by H. influenza was lower than that reported for other countries. ResumoObjetivos: 1) Determinar a prevalência do Streptococcus pneumoniae, Haemophilus influenzae e Moraxella catarrhalis nas efusões de orelha média de crianças com otite média com efusão que foram submetidas à miringotomia; 2) comparar os resultados obtidos por cultura e PCR; e 3) determinar o perfil de resistência à penicilina dos germes isolados.Métodos: Analisaram-se 128 amostras de efusões de orelha média de 75 crianças entre 11 meses e 10 anos de idade (média = 34,7 meses). Pacientes com otite média recorrente tinham efusão documentada por > 6 semanas e aqueles com otite média com efusão crônica, por > 3 meses. Os pacientes não tinham sinais de otite média aguda ou infecção do trato respiratório e não estavam sob antibioticoterapia no momento do pro...

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Section: Discussionunclassified

Prevalence of bacteria in children with otitis media with effusion

Pereira¹,

Pereira²,

Cantarelli³

et al. 2004

J Pediatr (Rio J)

View full text Add to dashboard Cite

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“…17 Animal studies have shed some light on the use of PCR for the investigation of the infectious etiology of middle ear infections. By using the OM model in chinchillas, Post et al 18 demonstrated that purified bacterial DNA and the DNA of heat-inactivated bacteria did not remain amplifiable for more than three days after inoculation. In addition, antibiotic-sensitive bacteria could not be cultured after the third day of treatment, but the DNA remained amplifiable for three weeks.…”

Section: Discussionmentioning

confidence: 99%

Prevalência de bactérias em crianças com otite média com efusão

Pereira¹,

Pereira²,

Cantarelli³

et al. 2004

J. Pediatr. (Rio J.)

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Objectives: 1) To determine the prevalence of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in middle ear effusions of children with otitis media with effusion undergoing myringotomy; 2) to compare the results obtained by culture and PCR; and 3) to determine the susceptibility of bacterial isolates to penicillin. Methods:We analyzed 128 middle ear effusion specimens from 75 children (age = 11 months to 10 years; mean = 34.7 months). Patients with recurrent otitis media had documented middle ear effusion for = 6 weeks, and chronic otitis media with effusion for = 3 months. The patients had no signs of acute otitis media or respiratory tract infection and were not on antibiotic therapy. Aspiration was done through tympanocentesis with an Alden-Senturia trap. Bacteriological studies were initiated less than 15 minutes after specimen collection. Part of the sample was stored at -20 o C for later multiplex PCR analysis. Statistical analysis employed McNemar s χ 2 test.Results: Bacteria were cultured in 32 (25.1%) out of 128 samples and the pathogens under investigation were found in 25 (19.6%). PCR was positive for bacteria in 73 (57.0%) specimens: 50 (39.1%) for H. influenzae, 16 (12.5%) for S. pneumoniae, and 13 (10.2%) for M. catarrhalis. All the culture-positive samples were PCR-positive, but 48 (65.7%) of the PCR-positive specimens were culture-negative. PCR was significantly more sensitive than culture (p < 0.01) to identify bacteria. Resistance to penicillin was as follows: M. catarrhalis = 100%; S. pneumoniae = 62.5% and H. influenzae = 23% of the isolates. Conclusions:The prevalence of bacteria in otitis media with effusion in a group of Brazilian children was similar to that reported for other countries. H. influenzae was the most frequent microorganism observed. This suggests that bacteria may play a role in the pathogenesis of otitis media with effusion. In addition, PCR was more sensitive to detect bacteria in middle ear effusion as compared to conventional culture methods. Penicillin resistance was similar to that reported for other countries for pneumococci and moraxella, but beta-lactamase production by H. influenza was lower than that reported for other countries.J Pediatr (Rio J). 2004;80(1):41-8: Otitis media, otitis media with effusion/microbiology, ear, middle/ microbiology, PCR, prevalence studies, child.

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“…The finding of bacterial mRNA is prima facie evidence that viable bacteria were present in the joint at the time of explant since bacterial mRNA has a half-life that is typically measured in seconds and/ or minutes and would therefore not be detectable if the bacteria were not viable 21 . While we believe that the combination of confocal microscopy and reverse transcriptase-polymerase chain reaction is a powerful tool for the direct detection of biofilms, the approach is not without limitations.…”

Section: Reverse Transcriptase-polymerase Chain Reaction and Polymeramentioning

confidence: 99%