THE USE OF MOLECULAR APPROACH (RISA) FOR DISCRIMINATION OF BACTERIAL SPECIES OF
THE ACTIVE BENIGN CHRONIC SUPPURATIVE OTITIS MEDIA
Anton Christanto, Soepomo Soekardono, Agus Surono, Novi Primadewi,
Roikhan Harowi.
Department of Otolaryngology Head and Neck Surgery, Faculty of Medicine,
University of Gadjah Mada/DR. Sarjito Hospital, Yogyakarta
ABSTRACT
Background: A bacterial ear infection, such as active benign chronic
suppurative otitis media (ABCSOM) is a potentially serious disorder and
requires topical antibiotic treatment. Traditionally, the identification and
enumeration of bacterial ear infection has depended entirely upon pure-culture
techniques. Observations of colony/cell morphology and biochemistry test
sometime do not give enough information to discriminate bacteria species
resulting unsatisfied treatment in some case. The intergenic spacers (IGS)
between the 16S and 23S rRNA genetic loci are frequently used in PCR
fingerprinting to discriminate bacterial strains at the species and
intraspecies levels.
Objective: In this study we investigated the molecular nature of polymorphisms
in IGS-PCR fingerprinting of bacteria species isolated from the active benign
chronic suppurative otitis media
Methods: Middle ear fluid discharge samples were collected from 5 individuals
of ABCSOM using sterile syringe equipped with intravenous catheter. Bacteria
were isolated from the fluid discharge samples by using plate method on blood
agar media. In this study we compared the traditional and molecular approach
(ribosomal intergenic spacer analysis-RISA) to discriminate bacterial species
obtained from fluid discharge samples. The RISA was performed by PCR
amplification of 16S-23S ribosomal DNA.
Results: The results showed the use of molecular approach revealed several
bacterial species isolated from fluid discharge samples, whereas the use of
traditional approach revealed only 2 type bacteria species in all samples.
Conclusion: The use of RISA for discrimination of bacterial species from active
benign chronic suppurative otitis media is robust method. It might be able to
discriminate bacterial ear pathogen at the species and intraspecies levels.
Key words: ABCSOM, ribosomal intergenic spacer analysis, bacterial species
discrimination
dr. HM Anton Christanto
Bag/SMF THT RSUP Dr Sardjito-FK UGM
Jl. Kesehatan no 1 Yogyakarta - Indonesia 55284
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