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Sowmiya M.,Kamal Nayan Bajaj Institute for Research in Vision and Ophthalmology | Sowmiya M.,Birla Institute of Technology and Science | Malathi J.,Kamal Nayan Bajaj Institute for Research in Vision and Ophthalmology | Swarnali S.,Sankara Nethralaya | And 3 more authors.
Indian Journal of Medical Research | Year: 2015

Background & objectives: There are only a few reports available on characterization of Propionibacterium acnes isolated from various ocular clinical specimens. We undertook this study to evaluate the role of P. acnes in ocular infections and biofilm production, and also do the phylogenetic analysis of the bacilli. Methods: One hundred isolates of P. acnes collected prospectively from ocular clinical specimens at a tertiary care eye hospital between January 2010 and December 2011, were studied for their association with various ocular disease conditions. The isolates were also subjected to genotyping and phylogenetic analysis, and were also tested for their ability to produce biofilms. Results: Among preoperative conjunctival swabs, P. acnes was a probably significant pathogen in one case; a possibly significant pathogen in two cases. In other clinical conditions, 13 per cent isolates were probably significant pathogens and 38 per cent as possibly significant pathogens. The analysis of 16S rRNA gene revealed four different phylogenies whereas analysis of recA gene showed two phylogenies confirming that recA gene was more reliable than 16S rRNA with less sequence variation. Results of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) had 100 per cent concordance with phylogenetic results. No association was seen between P. acnes subtypes and biofilm production. Interpretation & conclusions: RecA gene phylogenetic studies revealed two different phylogenies. RFLP technique was found to be cost-effective with high sensitivity and specificity in phylogenetic analysis. No association between P. acnes subtypes and pathogenetic ability was observed. Biofilm producing isolates showed increased antibiotic resistance compared with non-biofilm producing isolates. © 2015, Indian Council of Medical Research. All rights reserved. Source

Aarthi P.,Kamal Nayan Bajaj Institute for Research in Vision and Ophthalmology | Bagyalakshmi R.,Kamal Nayan Bajaj Institute for Research in Vision and Ophthalmology | Therese K.L.,Kamal Nayan Bajaj Institute for Research in Vision and Ophthalmology | Malathi J.,Kamal Nayan Bajaj Institute for Research in Vision and Ophthalmology | And 2 more authors.
Current Eye Research | Year: 2012

Purpose: To develop RNA based assay - reverse transcriptase polymerase chain reaction (RT-PCR) to detect viable bacteria in intraocular specimens obtained from patients with infectious endophthalmitis. Materials and methods: Thirty-five intraocular specimens (19 vitreous fluid and 16 aqueous humor) collected from patients with typical infectious endophthalmitis were subjected to conventional and molecular microbiological investigations. Culture negative, eubacterial genome PCR positive intraocular specimens were subjected to denaturing high performance liquid chromatography (dHPLC) for separation of mixed genomes and subsequently identified by PCR based DNA sequencing. In parallel, RT-PCR was performed to detect the presence of viable bacteria in intraocular specimens. Results: Among 35 intraocular specimens, single bacterial genome was detected in 9 (25.7%) and two or more genomes in 26 (74.28%) intraocular specimens. Eubacterial genome was detected by RT-PCR in 29 (82.85%) specimens. PCR based dHPLC followed by PCR based DNA sequencing revealed the presence of 65 bacterial genomes in 35 intraocular specimens. Five novel genera namely Terrabacter species, Facklamia species, Xylella fastidiosa, Duganella species and Synechococcus species were detected. Conclusion: RT-PCR serves as a rapid and reliable tool to detect viable bacteria causing endophthalmitis. © 2012 Informa Healthcare USA, Inc. Source

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