Eubacterial PCR for Bacterial Detection and Identification in 100 Acute Postcataract Surgery Endophthalmitis

• Published in: Investigative ophthalmology & visual science 2008-05, Vol.49 (5), p.1971-1978
• Main Authors: Chiquet, Christophe, Cornut, Pierre-Loic, Benito, Yvonne, Thuret, Gilles, Maurin, Max, Lafontaine, Pierre-Olivier, Pechinot, Andre, Palombi, Karine, Lina, Gerard, Bron, Alain, Denis, Philippe, Carricajo, Anne, Creuzot, Catherine, Romanet, Jean-Paul, Vandenesch, Francois, French Institutional Endophthalmitis Study (FRIENDS) Group
• Format: Article
• Language: English
• Subjects: Acute Disease; Aged; Aqueous Humor; Aqueous Humor - microbiology; Bacteria; Bacteria - genetics; Bacteria - isolation & purification; Bacteriological Techniques; Biochemistry, Molecular Biology; Biological and medical sciences; DNA, Bacterial; DNA, Bacterial - analysis; Endophthalmitis; Endophthalmitis - microbiology; Eye and associated structures. Visual pathways and centers. Vision; Eye Infections, Bacterial; Eye Infections, Bacterial - microbiology; Female; Fundamental and applied biological sciences. Psychology; Humans; Life Sciences; Male; Medical sciences; Ophthalmology; Phacoemulsification; Polymerase Chain Reaction; Polymerase Chain Reaction - methods; Postoperative Complications; Prospective Studies; RNA, Ribosomal, 16S; RNA, Ribosomal, 16S - genetics; Sequence Analysis, DNA; Vertebrates: nervous system and sense organs; Vitreous Body; Vitreous Body - microbiology
• ISSN: 0146-0404; 1552-5783; 1552-5783
• DOI: 10.1167/iovs.07-1377

To evaluate eubacterial PCR compared with conventional cultures for detection and identification of bacterial agents in ocular samples from patients with acute postcataract endophthalmitis. Broad-range eubacterial PCR amplification was used, followed by direct DNA sequencing in ocular samples (aqueous humor, vitreous samples from tap or vitrectomy) from 100 consecutive patients presenting with acute postcataract endophthalmitis. Bacterial cultures were performed on the same ocular samples by using traditional methods (brain-heart infusion broth). At the time of admission, the detection rate was not significantly different between cultures and PCR (38.2% for cultures versus 34.6% for PCR in aqueous humor samples; 54% versus 57% in vitreous from a vitreous tap). In contrast, in the vitreous obtained from vitrectomy, after intravitreous injection of antibiotics, PCR detected bacteria in 70% of the cases, compared with 9% in cultures. By combining PCR and cultures, bacterial identification was obtained in 47% of aqueous humor samples at admission, in 68% of vitreous samples from a vitreous tap at admission, and in 72% of vitreous samples from pars plana vitrectomy. Gram-positive bacteria predominated (94.3%). The concordance between cultures and PCR was 100%. The contamination rate was 2%. Cultures and eubacterial PCR are complementary techniques for bacterial identification in eyes with acute postcataract endophthalmitis. PCR technique was needed for identification of the involved microbial pathogen in 25% of all the cases. Eubacterial PCR is more effective than cultures in detecting bacteria in vitreous samples from patients with previous intravitreous administration of antibiotics.