oa Molecular Diagnosis and Vaccines - Diagnosis of bacterial endophthalmitis by polymerase chain reaction using universal primers for eubacterial genome



Endophthalmitis is a devastating ocular disease, the incidence of which has been reported as 0.1-0.3% after intraocular surgery and 2.0-3.0% after penetrating ocular trauma. Confirmation of the diagnosis of bacterial endophthalmitis is dependent onmicrobiological isolation of organisms. We evaluated polymerase chain reaction (PCR) in the diagnosis of bacterial endophthalmitis using vitreous fluid (VF) and aqueous humour (AH) specimens. Intraocular specimens from 50 (20 VF and 30 AH) cases with endophthalmitis and 20 (6 VF and 14AH) cases with non-infective disorders, were processed for microbiological investigations. Nested PCR directed at the 16S rDNA using universal primers for eubacterial genome was done. Of the 20 controls 2 (10%) were positive using eubacterial primer. On the other hand PCR for eubacterial genome showed 100% correlation with the 22 (44%) bacteriologically positive specimens. Eubacterial genome, was detected in 9 (18%) of 28 bacteriologically negative specimens. Among the 19 eubacterial PCR negative specimens, 8 were fungus positive Candida (5) and Aspergillus (3). By inclusion of PCR, microbiologically positive specimens increased from 46.5% to 75.8%. Application of PCR on AH was as sensitive as that on VF for the detection of eubacterial genome. It is concluded that PCR is a reliable tool for the diagnosis of bacterial endophthalmitis.


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