An application of outer membrane protein p6-specific enzyme-linked immunosorbent assay for detection of haemophilus influenzae in middle ear fluids and nasopharyngeal secretions

PLoS One. 2013 Aug 28;8(8):e71774. doi: 10.1371/journal.pone.0071774. eCollection 2013.

Abstract

An enzyme-linked immunosorbent assay specific to outer membrane protein P6 (P6-ELISA) was applied for detecting Haemophilus influenzae in middle ear fluids (MEFs) from acute otitis media (AOM) patients and in nasopharyngeal secretions (NPSs) from acute rhinosinusitis patients. P6-ELISA had a sensitivity of 83.3% for MEFs and 71.5% for NPSs and a specificity of 85.6% for MEFs and 92.5% for NPSs, respectively. Real-time PCR exhibited significant differences in the number of ompP1 gene copies among samples determined by P6-ELISA to be positive and negative for H. influenzae. However, because the P6-ELISA test has the reactivity in Haemophilus species include two commensals H. haemolyticus and H. parainfluenzae, it is thus a weak method in order to detect only NTHi correctly. Consequently, diagnosis using the P6-ELISA should be based on an overall evaluation, including the results of other related examinations and clinical symptoms to prevent misleading conclusions in clinical setting.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acute Disease
  • Adolescent
  • Adult
  • Aged
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / metabolism*
  • Child
  • Child, Preschool
  • Ear, Middle / microbiology
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Haemophilus Infections / diagnosis*
  • Haemophilus Infections / microbiology
  • Haemophilus Vaccines / genetics
  • Haemophilus Vaccines / metabolism*
  • Haemophilus influenzae / genetics
  • Haemophilus influenzae / metabolism*
  • Humans
  • Infant
  • Male
  • Middle Aged
  • Nasopharynx / microbiology
  • Otitis Media / diagnosis*
  • Otitis Media / microbiology
  • Rhinitis / diagnosis*
  • Rhinitis / microbiology
  • Sensitivity and Specificity
  • Sinusitis / diagnosis*
  • Sinusitis / microbiology
  • Young Adult

Substances

  • Bacterial Outer Membrane Proteins
  • Haemophilus Vaccines
  • P6 outer membrane protein, Haemophilus

Grants and funding

This study was funded by Otsuka Pharmaceutical Co. Ltd., Tokyo Japan, whose role, though the employment of Hisayo Masuda, Hideaki Tanaka, and Yumi Tanaka, involved technical assistance for performing this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.