Identification of pathgenic bacteria in aqueous and vitreous of endophthalmitis by 16S rDNA sequencing technique

• DOI: 10.3760/cma.j.issn.2095-0160.2016.10.004
• Publication Date: 2016-10-10
• Authors: Baoxia Yang; Hong Li; Fanfang Kong
• Keywords: Hypervariable region

Background Endophthalmitis is a serious complication of intraocular surgery.Conventional identification methods for bacteria are becterial culture and smear method, but these laboratory tests spend long time have low positive rates.16S rDNA the bacterial chromosome encoding ribosomal RNA sequences, it determined that 16S sequencing has high specificity bacteria.

Objective This study was to identify infectious from aqueous humor or vitreous body in eyes with endophthalmitis by technique, investigate diagnosis efficency technique on endophthalmitis.

Methods Anterior chamber fluid (0.1-0.2 ml) (0.5-1.0 specimens were collected 5 patients Qingdao Eye Hospital June December 2015 used throughput sequencing, smear, respectively.Bacteria DNA extracted specimen D3096-01 trace kit amplification V3-V4 region rRNA gene hypervariable all microbes samples MiSeq Illumina Sequencing Platform.Then bioinformatic analysis including taxonomy, abundance alpha diversity performed.Nuclease-free water 50 μl centrifuge tube as control.

Results Five collected, results exhibited examination, Gram bacilli trumatic eye negative filtering bleb eye, negative.16S squencing showed outcomes samples.The abundent nacteral genuses Staphylococcus (65.28%), Streptococcus (18.90%) Pseudomonas (12.76%) eye; major components sample (53.68%), Acinetobacter (8.62%) Limnobacter (5.96%) acute occurring at 2 days following cataract surgery; Moraxella (88.89%) (9.52%); later-onset (84.63%) 1 day after surgery (97.89%).

Conclusions A distinct advantage found indentification pathogenic due its rate comparison method.

Key words: Endophthalmitis/microbiology; Aqueous humor/microbiology; Vitreous body/microbiology; DNA, bacterial/analysis; Sequence analysis, DNA; 16S