Had keratitis and endophthalmitis by amplification and subsequentĭirect sequencing of 16S rDNA. 4 detected and identifiedīacteria in corneal scrapings and in vitreous samples of patients who In vitreous samples of patients who had endophthalmitis. 2 used eubacterial primers and Propionibacterium-specific primers to detect bacterial DNA Pathogens is still at its beginning and, except in a few studies, is In ophthalmology, the 16S rDNA-based identification of The comparison ofĪmplified and sequenced 16S rDNA sequences with sequences of knownīacteria in 16S rDNA databases facilitates a subsequent phylogenetic Targeted to highly conserved regions are applied. Possible without prior cultivation when broad-range PCR primers The amplification of 16S rDNA of any bacterial species is Molecular approaches to the identification of bacteria show promising Methods for the detection and identification of monomicrobial as wellĪs polymicrobial ocular infections of bacteria that might not be 16S rDNA sequence analyses and DGGE fingerprinting are appropriate No sample positive by culture gave negative results by PCR.Ĭonclusions. In total, 45% of the 60 analyzedĬonjunctival swabs were PCR positive, whereas only 22% were culture ( n = 1), Klebsiella oxytoca ( n = 1), and Pseudomonas aeruginosa ( n = 1). ( n = 9), Corynebacteria ( n = 3), Staphylococcus aureus ( n = 1), Streptococcus sp. Sequences of Pantoea and Enterobacter ( n = 1), Kingella and Neisseria ( n = 1), Serratia and Aranicola ( n = 1), and Leuconostoc and Weissella ( n = 2), respectively.Ĭulture was only positive for coagulase-negative staphylococci They had highest sequence similarities both to Four sequences could not be identified to Pseudomonas ( n = 3), Proteus ( n = 1), and Brevundimonas ( n = 1). Streptococcus ( n = 6), Bacillus ( n = 2) , Theįollowing genera ( n is number of samples) were detected: Staphylococcus ( n = 8), Corynebacterium ( n = 7) ,
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Monomicrobial and 11 samples showed polymicrobial infections. Purulent conjunctivitis eyes and from 2 of 31 investigated swabs takenįrom nonpurulent conjunctivitis eyes. 16S rDNA could be amplified from 25 of 29 investigated swabs taken from Furthermore, the results were compared with results Sequences were compared with sequences of known bacteria listed in theĮMBL database. RDNA clone libraries were constructed and clones were screened by DGGE. Identification, DGGE bands were excised and directly sequenced, or 16S Fragments ofĢ00 bp, spanning the V3 region of the eubacterial 16S rDNA, wereĪmplified by polymerase chain reaction (PCR) and separated byĭenaturing gradient gel electrophoresis (DGGE). From 60 human conjunctivae (29 with purulent and 31 with nonpurulentĬonjunctivitis), swabs were taken and DNA was extracted. Identification of multiple bacteria from the ocular environment, whichĬan be applied supplementarily to cultivation in cases of severe Establishment of a new molecular biology technique for the