Accuracy of real-time PCR, Gram stain and culture for Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae meningitis diagnosis

BMC Infectious Diseases

Joice N Reis⁵  Stacey W Martin⁴  Albert I Ko¹  Leonard W Mayer⁴  Thomas A Clark⁴  Brian D Plikaytis⁴  Mitermayer G Reis²  Katia Salgado²  Mariela C Leite⁵  Tainara Q Oliveira⁵  Jailton Azevedo²  MariadaGloriaS Carvalho⁴  Brian H Harcourt⁴  Soraia M Cordeiro²  Henry M Wu³ 

[1] Weill Medical College of Cornell University, 1300 York Avenue, New York, NY, 10065, USA;Oswaldo Cruz Foundation, Rua Waldemar Falcão 121, Candeal, Salvador Bahia, 40296-710, Brazil;Present address: 550 Peachtree Street NE, MOT 7, Atlanta, GA, 30308, USA;Division of Bacterial Diseases, National Center for Immunization and Respiratory Diseases, CDC, 1600 Clifton Road, Atlanta, GA, 30333, USA;Federal University of Bahia, Salvador, Bahia, Brazil

关键词: Haemophilus influenzae;    Neisseria meningitidis;    Streptococcus pneumoniae;    Real-time PCR;    Diagnostic test evaluation;    Bacterial meningitis;   

Others  :  1158554
DOI  :  10.1186/1471-2334-13-26

 received in 2012-09-06, accepted in 2013-01-17,  发布年份 2013

【 摘 要 】

Background

Although cerebrospinal fluid (CSF) culture is the diagnostic reference standard for bacterial meningitis, its sensitivity is limited, particularly when antibiotics were previously administered. CSF Gram staining and real-time PCR are theoretically less affected by antibiotics; however, it is difficult to evaluate these tests with an imperfect reference standard.

Methods and findings

CSF from patients with suspected meningitis from Salvador, Brazil were tested with culture, Gram stain, and real-time PCR using S. pneumoniae, N. meningitidis, and H. influenzae specific primers and probes. An antibiotic detection disk bioassay was used to test for the presence of antibiotic activity in CSF. The diagnostic accuracy of tests were evaluated using multiple methods, including direct evaluation of Gram stain and real-time PCR against CSF culture, evaluation of real-time PCR against a composite reference standard, and latent class analysis modeling to evaluate all three tests simultaneously.

Results

Among 451 CSF specimens, 80 (17.7%) had culture isolation of one of the three pathogens (40 S. pneumoniae, 36 N. meningitidis, and 4 H. influenzae), and 113 (25.1%) were real-time PCR positive (51 S. pneumoniae, 57 N. meningitidis, and 5 H. influenzae). Compared to culture, real-time PCR sensitivity and specificity were 95.0% and 90.0%, respectively. In a latent class analysis model, the sensitivity and specificity estimates were: culture, 81.3% and 99.7%; Gram stain, 98.2% and 98.7%; and real-time PCR, 95.7% and 94.3%, respectively. Gram stain and real-time PCR sensitivity did not change significantly when there was antibiotic activity in the CSF.

Conclusion

Real-time PCR and Gram stain were highly accurate in diagnosing meningitis caused by S. pneumoniae, N. meningitidis, and H. influenzae, though there were few cases of H. influenzae. Furthermore, real-time PCR and Gram staining were less affected by antibiotic presence and might be useful when antibiotics were previously administered. Gram staining, which is inexpensive and commonly available, should be encouraged in all clinical settings.

【 授权许可】

   
2013 Wu et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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