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

Muneki Hotomi; Akihisa Togawa; Masamitsu Kono; Gen Sugita; Rinya Sugita; Yutaka Fujimaki; Yosuke Kamide; Akihiro Uchizono; Keiko Kanesada; Shoichi Sawada; Naohiro Okitsu; Hisayo Masuda; Hideaki Tanaka; Yumi Tanaka; Noboru Yamanaka

doi:10.1371/journal.pone.0071774

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.

Authors

Muneki Hotomi¹, Akihisa Togawa, Masamitsu Kono, Gen Sugita, Rinya Sugita, Yutaka Fujimaki, Yosuke Kamide, Akihiro Uchizono, Keiko Kanesada, Shoichi Sawada, Naohiro Okitsu, Hisayo Masuda, Hideaki Tanaka, Yumi Tanaka, Noboru Yamanaka

Affiliation

¹ Department of Otolaryngology-Head and Neck Surgery, Wakayama Medical University, Wakayama, Japan.

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.