- 标题
- 摘要
- 关键词
- 实验方案
- 产品
-
Peptide Nucleic Acid–Fluorescence In Situ Hybridization for Detection of Staphylococci From Endophthalmitis Isolates: A Proof-of-Concept Study
摘要： PURPOSE. Rapid identi?cation of pathogens causing endophthalmitis may improve treatment outcomes through early administration of species-speci?c medication. The current study reports a new molecular application of peptide nucleic acid–?uorescence in situ hybridization (PNA-FISH) with Staphylococcus-speci?c molecular PNA probes for the potential rapid detection of common pathogens causing endophthalmitis. METHODS. An experimental study was designed to evaluate the proof of concept at the microbiology laboratory of the Bascom Palmer Eye Institute. Stored culture-positive staphylococci endophthalmitis isolates obtained from prior vitreous samples (n ? 15), along with broth as negative controls (n ? 5) were used. Inoculum was prepared to a ?nal concentration of 1 3 105 colony-forming units/mL to ensure that the isolates were viable. Smears of samples were ?xed and hybridized using QuickFISH protocol with probes for Staphylococcus. RESULTS. With PNA-FISH technique, Staphylococcus aureus was identi?ed in 9 of 10 samples and coagulase-negative staphylococci were identi?ed in 10 of 10 samples. Detection time was 20 minutes. CONCLUSIONS. This study serves a proof of concept using a new microbial detection system with FISH probes, and may have the potential for clinical use in the rapid and accurate identi?cation of isolates from patients with endophthalmitis.
关键词： peptide nucleic acid (PNA),endophthalmitis,rapid identification,fluorescence in situ hybridization (FISH)
更新于2025-09-23 15:22:29
-
Comparing consistency of clear corneal incisions using a traditional diamond keratome and a newly designed diamond keratome
摘要： Purpose: To compare the consistency of incision architecture utilizing a traditional diamond keratome and a newly designed diamond keratome. Methods: We used a traditional diamond keratome and newly designed diamond keratome to create clear corneal incisions in human cadaveric donor eyes. Three surgeons with varying levels of experience made 30 incisions with each keratome; and the wound architecture was measured including incision lengths, epithelial and endothelial widths, and the central epithelial incision’s deviation from a straight line entrance. Results: The mean absolute difference in right and left incision lengths (traditional: 0.182±0.158 mm and new 0.088±0.077 mm [P<0.003]), mean absolute difference in epithelial and endothelial incision width (traditional: 0.181±0.144 mm and new 0.080±0.092 mm [P<0.002]), endothelial incision central deviation from a straight line (traditional: 0.128±0.242 mm and new -0.046±0.124 mm [P<0.001]) were all significantly more consistent with the newly designed diamond keratome than with the traditional diamond keratome. Conclusion: The newly designed diamond keratome creates a more consistent clear corneal incision than a traditional diamond keratome across a variety of surgical skill levels and does not require advanced levels of training to achieve this consistency.
关键词： keratome,postoperative endophthalmitis,corneal incision
更新于2025-09-23 15:22:29
-
Blood–Retinal Barrier Compromise and Endogenous <i>Staphylococcus aureus</i> Endophthalmitis
摘要： PURPOSE. To test the hypothesis that blood–retinal barrier compromise is associated with the development of endogenous Staphylococcus aureus endophthalmitis. METHODS. To compromise the blood–retinal barrier in vivo, streptozotocin-induced diabetes was induced in C57BL/6J mice for 1, 3, or 5 months. Diabetic and age-matched nondiabetic mice were intravenously injected with 108 colony-forming units (cfu) of S. aureus, a common cause of endogenous endophthalmitis in diabetics. After 4 days post infection, electroretinography, histology, and bacterial counts were performed. Staphylococcus aureus–induced alterations in in vitro retinal pigment epithelial (RPE) cell barrier structure and function were assessed by anti–ZO-1 immunohistochemistry, FITC-dextran conjugate diffusion, and bacterial transmigration assays. RESULTS. We observed one bilateral infection in a control, nondiabetic animal (mean ? 1.54 3 103 6 1.78 3 102 cfu/eye, 7% incidence). Among the 1-month diabetic mice, we observed culture-con?rmed unilateral infections in two animals (mean ? 5.54 3 102 6 7.09 3 102 cfu/ eye, 12% incidence). Among the 3-month diabetic mice, infections were observed in 11 animals, three with bilateral infections (mean ? 2.67 3 102 6 2.49 3 102 cfu/eye, 58% incidence). Among the 5-month diabetic mice, we observed infections in ?ve animals (mean ? 7.88 3 102 6 1.08 3 103 cfu/eye, 33% incidence). In vitro, S. aureus infection reduced ZO-1 immunostaining and disrupted the barrier function of cultured RPE cells, resulting in diffusion of ?uorophore-conjugated dextrans and transmigration of live bacteria across a permeabilized RPE barrier. CONCLUSIONS. Taken together, these results indicated that S. aureus is capable of inducing blood–retinal barrier permeability and causing endogenous bacterial endophthalmitis in normal and diabetic animals.
关键词： Staphylococcus aureus,endogenous endophthalmitis,blood–retinal barrier,bacteria,diabetes,retinal pigment epithelium
更新于2025-09-23 15:21:01
-
Rapid pathogen identification and antimicrobial susceptibility testing in in vitro endophthalmitis with matrix assisted laser desorption-ionization Time-of-Flight Mass Spectrometry and VITEK 2 without prior culture
摘要： Prompt clinical diagnosis and initiation of treatment are critical in the management of infectious endophthalmitis. Current methods used to identify causative agents of infectious endophthalmitis are mostly inefficient, owing to suboptimal sensitivity, length, and cost. Matrix Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) can be used to rapidly identity pathogens without a need for culture. Similarly, automated antimicrobial susceptibility test systems (AST, VITEK 2) provide accurate antimicrobial susceptibility profiles. In this proof-of-concept study, we apply these technologies for the direct identification and characterization of pathogens in vitreous samples, without culture, as an in vitro model of infectious endophthalmitis.
关键词： endophthalmitis,pathogen identification,MALDI-TOF MS,antimicrobial susceptibility testing,VITEK 2
更新于2025-09-12 10:27:22
-
LONG-TERM VISUAL OUTCOMES AND CLINICAL FEATURES AFTER ANTI–VASCULAR ENDOTHELIAL GROWTH FACTOR INJECTION–RELATED ENDOPHTHALMITIS
摘要： Purpose: To determine long-term visual outcomes in patients who developed endophthalmitis after intravitreal anti–vascular endothelial growth factor injections and to correlate visual outcomes with clinical features. Methods: This is a retrospective, multicenter, consecutive case series of patients diagnosed with anti–vascular endothelial growth factor injection–related endophthalmitis who were treated at Mid Atlantic Retina, the Retina Service of Wills Eye Hospital, Philadelphia, PA, and the University of Southern California Roski Eye Institute, Los Angeles, CA. Patients were included if they had at least 1 year of follow-up. Primary outcome was to evaluate long-term visual outcomes up to 5 years of follow-up. The secondary outcome was to determine clinical features (e.g., culture results) that may predict long-term visual acuity outcomes. Results: A total of 56 cases of endophthalmitis from 168,247 anti–vascular endothelial growth factor injections were identi?ed (0.033%, 1/3,004 injections), from which 51 eyes met inclusion criteria. Mean follow-up period was 3.3 years (median 4 years; range 1–5 years). A total of 24 patients (47%) reached a maximum ?nal follow-up of 5 years. Mean Snellen visual acuity at the causative injection visit was 20/102 and decreased to counting ?ngers at diagnosis (P , 0.001). At 6-month follow-up, mean visual acuity improved to 20/644 (P , 0.001) and remained stable up to 5 years (20/480, P = 0.003) follow-up compared with diagnosis. At the ?nal follow-up, 20 eyes had visual acuity that returned to within one line of baseline visual acuity (visual recovery group), whereas 31 patients’ visual acuity was at least one line worse than initial visual acuity (visual deterioration group). The cultures for the visual recovery group were more likely to grow coagulase-negative Staphylococcus, whereas the visual deterioration group primarily grew Streptococcus species, Staphylococcus aureus, and Enterococcus faecalis (P = 0.002, comparing organisms isolated in the visual recovery and deterioration group). Conclusion: Visual outcomes after anti–vascular endothelial growth factor injection– related endophthalmitis seem to reach peak improvement by 6 months and remain stable up to a median of 4-year follow-up. Patients who develop culture-negative endophthalmitis or endophthalmitis secondary to coagulase-negative Staphylococcus are more likely to regain baseline visual acuity compared with cases secondary to Streptococcus species.
关键词： bevacizumab,anti-vascular endothelial growth factor,diabetic macular edema,intravitreal injection,endophthalmitis,a?ibercept,long-term,ranibizumab,neovascular age-related macular degeneration,retinal vein occlusion
更新于2025-09-10 09:29:36
-
Comparison of microbiology and visual outcomes of patients undergoing small-gauge and 20-gauge vitrectomy for endophthalmitis
摘要： Background: The role of pars plana vitrectomy (PPV) for endophthalmitis has evolved over recent decades but the literature is lacking on comparisons between small-gauge and 20-gauge vitrectomy. Objective: To evaluate evolving etiological and microbiological trends in patients undergoing vitrectomy for endophthalmitis and to compare culture-positive rates and visual outcomes between small-gauge (23- and 25-gauge) and 20-gauge instrumentation during vitrectomy for endophthalmitis. Methods: Ten-year retrospective comparative case series and prospective laboratory in vitro testing. Tertiary care academic referral center. Patients who underwent PPV for endophthalmitis between 2003 and 2013. Vitreous biopsies were obtained in all cases. The effect of vitrectomy gauge (20-, 23-, and 25-gauge) and vitreous cutting rate (1,500 and 5,000 cuts per minute) on the viability of bacterial culture was evaluated in an in vitro prospective laboratory investigation. Main outcome measures: Comparison of etiology, microbiology culture-positive rates, and visual outcomes between small-gauge and 20-gauge instrumentation in patients undergoing PPV for infectious endophthalmitis. Results: A total of 61 cases of vitrectomy for endophthalmitis were identified over a 10-year period; of these, 34 were treated with small-gauge (23- and 25-gauge) vitrectomy and 27 were treated with 20-gauge vitrectomy. In the small-gauge group, 12 cases (35.3%) yielded culture-positive results versus 20 cases (74.1%) with culture positivity in the 20-gauge cohort (P=0.002). The most common cause of endophthalmitis was cataract surgery and the most frequently identified organism was coagulase-negative Staphylococci in both groups. There was no significant difference in mean postoperative visual acuities between groups (P=0.33). Etiological trends indicate an increase in endophthalmitis due to intravitreal injection in the small-gauge group (n=9) compared to the 20-gauge group (n=3) (P=0.001). In vitro laboratory testing revealed no significant difference in rates of culture growth for different vitrectomy gauge sizes or vitreous cutting speeds. Conclusion and relevance: Small-gauge vitrectomy for endophthalmitis yields final visual outcomes comparable to 20-gauge instrumentation. A significant difference in culture-positive rates was observed between small-gauge and 20-gauge instrumentation for vitrectomy in endophthalmitis; however, laboratory testing indicates this is not related to either vitreous gauge size or cutter speed. Intravitreal injections are emerging as a common etiology of vitrectomy for endophthalmitis.
关键词： vitrectomy,vitreous biopsy,endophthalmitis
更新于2025-09-04 15:30:14
-
Acute subretinal abscess in <i>Staphylococcus aureus</i> septicaemia with endophthalmitis showcased by multimodal retinal imaging and with 2-year follow-up
摘要： A middle-aged man presented to emergency services with central vision loss in the setting of flu-like illness with fever. A striking subfoveal abscess was observed in the right fundus. Focal acute chorioretinal inflammation was noted in the asymptomatic fellow eye. Staphylococcus aureus septicaemia was subsequently diagnosed. He presented with undiagnosed HIV infection and latent syphilis. Serial high-definition multimodal retinal imaging showcased resolution of the dome-shaped subretinal abscess following treatment with intravenous flucloxacillin. A chorioretinal scar swiftly replaced the subfoveal abscess. Peripheral right vision and full left vision was retained. Vision loss due to endogenous endophthalmitis in systemic sepsis is an emergency requiring prompt multidisciplinary care. Sight and life are at risk—thus this is not a diagnosis to miss! Early recognition is paramount to health and in retaining vision. We briefly review relevant literature and portray how multimodal imaging guided response to treatment of acute subretinal abscess.
关键词： endophthalmitis,Staphylococcus aureus septicaemia,multimodal retinal imaging,HIV infection,acute subretinal abscess,latent syphilis
更新于2025-09-04 15:30:14