Identification of bacteria in culture negative and polymerase chain reaction (PCR) positive intraocular specimen from patients with infectious endopthalmitis.
A novel Denaturing High-Performance Liquid Chromatography (dHPLC)-based technique allows rapid high-resolution analysis of PCR products. We show the application of this PCR/dHPLC approach for direct detection and identification of bacterium from the Eubacterial PCR amplified products of aqueous and vitreous aspirates from patients with endopthalmitis and to differentially identify the culture negative cases and initiate appropriate therapy. The aim of this study is to identify culture negative PCR positive cases by the application of PCR based DNA sequencing. A total of 116 intraocular specimens were subjected for the study. Sixty-nine different bacteria were identified using dHPLC based DNA sequencing of which predominant ones were Gram-positive bacteria and cannot be cultured by conventional methods. Forty eight different bacteria detected in this study is being reported for the first time in infectious endopthalmitis.
L & T Microbiology Research Center, Sankara Nethralaya, 18, College Road, Chennai 600006, India.
Pasupathi
P
Aarthi
Rajagopal
R
Harini
Murali
M
Sowmiya
Jambulingam
J
Malathi
K Lily
KL
Therese
Hajib N
HN
Madhavan
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001431:Bacteriological Techniques / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015818:Eye Infections, Bacterial / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial