Random Amplified Polymorphic DNA Subtyping of Haemophilus influenzae from Middle Ear Effusion Fluid of Lebanese Patients with Otitis Media with Effusion

Abstract

Objective: In this study, we evaluated a Random
Amplified Polymorphic DNA (RAPD) method, in the
subtyping of H. influenzae in middle ear effusions
(MEE), obtained from 33 patients with otitis media with
effusion (OME) admitted to 3 medical centers in Beirut,
Lebanon.
Method: RAPD was initially evaluated on 15 H.
influenzae isolates using three 10 mer primers along
with a 21 mer primer.
Results: Primer 1 (10 mer) and Primer 2 (20 mer) were
the most discriminatory when used in conjunction and
were selected to be used on DNA lysates obtained from
33 MEE samples positive for H. influenzae by PCR.
Conclusion: Our data have shown that 4 RAPD patterns
were obtained on DNA of H. influenzae isolates and a
single RAPD pattern was seen on H. influenzae DNA
from MEE samples. This indicates that a single strain
may have been implicated in these infections. Studies
are underway to determine the prevalence of H.
influenzae as the etiology of otitis media with effusion
in Lebanon and subtype the organism to determine a
possible multi-strain involvement.
Key words: Otitis, PCR, subtyping

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