Utility of broad-range 16S rRNA PCR assay versus conventional methods for laboratory diagnosis of bacterial endophthalmitis in a tertiary care hospital
| Autor: | Deepanshi Mishra, Subrat Kumar Panda, Gita Satpathy, Nishat Hussain Ahmed, Rohan Chawla, Pradeep Venkatesh |
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| Rok vydání: | 2018 |
| Předmět: |
DNA
Bacterial Pcr assay Polymerase Chain Reaction Eye Infections Bacterial Microbiology Tertiary Care Centers 03 medical and health sciences Cellular and Molecular Neuroscience 0302 clinical medicine Endophthalmitis RNA Ribosomal 16S medicine Humans Prospective Studies Bacteriological Techniques biology Bacteria business.industry Ribosomal RNA medicine.disease Antimicrobial Isolation (microbiology) 16S ribosomal RNA biology.organism_classification Sensory Systems Ophthalmology GenBank 030221 ophthalmology & optometry business 030217 neurology & neurosurgery |
| Zdroj: | The British journal of ophthalmology. 103(1) |
| ISSN: | 1468-2079 |
| Popis: | BackgroundEndophthalmitis, a sight-threatening intraocular infection, can be of postsurgical, post-traumatic or endogenous origin. Laboratory diagnosis-based appropriate therapy can be vision-saving. Conventional culture-based laboratory diagnosis takes time and lacks sensitivity. In this study a broad-range PCR assay was assessed against conventional and automated culture methods in vitreous specimens for accurate microbiological diagnosis.AimsTo use broad-range PCR assay targeting 16S ribosomal RNA (rRNA) region of bacteria and to assess its performance vis-à-vis conventional and automated culture methods in the laboratory diagnosis of endophthalmitis.MethodsVitreous specimens from 195 patients with clinically diagnosed endophthalmitis were processed for classical and automated culture methods, antimicrobial sensitivity and broad-range PCR assay targeting 762 bp region of 16S rRNA followed by nucleotide sequencing by Sanger’s method. Causative agents were identified from the nucleotide sequences analysed against the GenBank database, and organisms were identified using the Clinical and Laboratory Standards Institute (CLSI) MM18A guidelines.ResultsBacteria could be detected from 127 (65.13%) of the 195 vitreous specimens by broad-range PCR assay; bacterial isolation was possible from 17 (8.7%) and 60 (30.76%) of these specimens by conventional and automated culture methods, respectively (pConclusionBroad-range PCR assay could provide definitive microbial diagnosis within 24 hours in significantly more patients (p |
| Databáze: | OpenAIRE |
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