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Updated: May 11, 2026
In Situ Detection of Bacteria within Paraffin-embedded Tissues Using a Digoxin-labeled DNA Probe Targeting 16S rRNA
Published on: May 21, 2015
Rapid detection of bacterial DNA in mastoid granulation tissue with nested-PCR technique
Peem Eiamprapai1, Yasufumi Matsumura, Harukazu Hiraumi
1Department of Otolaryngology, Faculty of Medicine, Mahasarakham University, Mahasarakham, Thailand. peemprot@gmail.com
Objective:
To detect bacterial DNA in mastoid granulation tissue from patients with chronic suppurative otitis media (CSOM).
Material And Method:
A two-step polymerase chain reaction (nested polymerase chain reaction) technique was employed. A 16s rRNA universal primer common to all bacteria was used as a bracket primer for the first step PCR reaction. Primers specific to P aeruginosa and S. aureus were then used as nested primers for the second step PCR. Products of this process were identified by DNA sequencing.
Results:
Among 15 clinical specimens collected, five showed positive bands specific to the species P aeruginosa, and 11 showed bands specific to the genus Staphylococcus. DNA sequencing showed 99.7 to 100% accuracy for target organisms in clinical specimens with a positive signal. The average time taken to conduct the PCR procedure was about four hours
Conclusion:
The nested PCR technique described worked well, even when the size of the mastoid granulation tissue was very small.
