RESUMO
PURPOSE: To study the risk factors, clinical features, and treatment outcomes of patients with culture-negative keratitis (CNK). METHODS: A retrospective data review of 933 patients with CNK was performed from January 2018 to December 2020. The variables such as the history of injury, visual acuity, slit-lamp findings with measurements of size and depth of ulcer, microbiological evaluation, duct patency, blood glucose levels, and treatment were considered, and clinical outcome was analyzed. RESULTS: Of the 933 patients with CNK, 763 (81.8%) were medically managed, with a mean treatment duration of 2.08 ± 1.7 weeks. Among them, 622 (66.7%) were both smear and culture-negative, and 311 (33.3%) showed only smear positivity. Smear-positive patients showed a positive correlation with the history of injury. A higher incidence of fungal growth on repeat culture was observed. Surgical interventions were done only in 18.2% of the patients; the rest were treated with topical medications alone. CONCLUSION: High clinical suspicion, differentiation of causative organisms based on clinical findings, and initiating empirical therapy with broad-spectrum antibiotics and antifungals improve the ultimate prognosis in patients with CNK, even though a standard protocol for empirical medical treatment may differ among institutions and surgeons based on their clinical experience and geographical variations.
Assuntos
Antibacterianos , Infecções Oculares Bacterianas , Infecções Oculares Fúngicas , Acuidade Visual , Humanos , Estudos Retrospectivos , Feminino , Masculino , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/tratamento farmacológico , Pessoa de Meia-Idade , Acuidade Visual/fisiologia , Adulto , Infecções Oculares Fúngicas/microbiologia , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/terapia , Antibacterianos/uso terapêutico , Bactérias/isolamento & purificação , Antifúngicos/uso terapêutico , Ceratite/microbiologia , Ceratite/diagnóstico , Ceratite/tratamento farmacológico , Fatores de Risco , Seguimentos , Fungos/isolamento & purificação , Córnea/microbiologia , Córnea/patologia , IdosoRESUMO
Importance: Microbial keratitis (MK) is a common cause of unilateral visual impairment, blindness, and eye loss in low-income and middle-income countries. There is an urgent need to develop and implement rapid and simple point-of-care diagnostics for MK to increase the likelihood of good outcomes. Objective: To evaluate the diagnostic performance of the Aspergillus-specific lateral-flow device (AspLFD) to identify Aspergillus species causing MK in corneal scrape and corneal swab samples of patients presenting with microbial keratitis. Design, Setting, and Participants: This diagnostic study was conducted between May 2022 and January 2023 at the corneal clinic of Aravind Eye Hospital in Madurai, Tamil Nadu, India. All study participants were recruited during their first presentation to the clinic. Patients aged 15 years or older met the eligibility criteria if they were attending their first appointment, had a corneal ulcer that was suggestive of a bacterial or fungal infection, and were about to undergo diagnostic scrape and culture. Main Outcomes and Measures: Sensitivity and specificity of the AspLFD with corneal samples collected from patients with MK. During routine diagnostic scraping, a minimally invasive corneal swab and an additional corneal scrape were collected and transferred to aliquots of sample buffer and analyzed by lateral-flow device (LFD) if the patient met the inclusion criteria. Photographs of devices were taken with a smartphone and analyzed using a ratiometric approach, which was developed for this study. The AspLFD results were compared with culture reports. Results: The 198 participants who met the inclusion criteria had a mean (range) age of 51 (15-85) years and included 126 males (63.6%). Overall, 35 of 198 participants with corneal scrape (17.7%) and 17 of 40 participants with swab samples (42.5%) had positive culture results for Aspergillus species. Ratiometric analysis results for the scrape samples found that the AspLFD achieved high sensitivity (0.89; 95% CI, 0.74-0.95), high negative predictive value (0.97; 95% CI, 0.94-0.99), low negative likelihood ratio (0.12; 95% CI, 0.05-0.30), and an accuracy of 0.94 (95% CI, 0.90-0.97). Ratiometric analysis results for the swab samples showed that the AspLFD had high sensitivity (0.94; 95% CI, 0.73-1.00), high negative predictive value (0.95; 95% CI, 0.76-1.00), low negative likelihood ratio (0.07; 95% CI, 0.01-0.48), and an accuracy of 0.88 (95% CI, 0.73-0.96). Conclusions and Relevance: Results of this diagnostic study suggest that AspLFD along with the ratiometric analysis of LFDs developed for this study has high diagnostic accuracy in identifying Aspergillus species from corneal scrapes and swabs. This technology is an important step toward the provision of point-of-care diagnostics for MK and could inform the clinical management strategy.
RESUMO
Purpose: To determine whether convolutional neural networks can detect morphological differences between images of microbiologically positive and negative corneal ulcers. Methods: A cross-sectional comparison of prospectively collected data consisting of bacterial and fungal cultures and smears from eyes with acute infectious keratitis at Aravind Eye Hospital. Two convolutional neural network architectures (DenseNet and MobileNet) were trained using images obtained from handheld cameras collected from culture-positive and negative images and smear-positive and -negative images. Each architecture was trained on two image sets: (1) one with labels assigned using only culture results and (2) one using culture and smear results. The outcome measure was area under the receiver operating characteristic curve for predicting whether an ulcer would be microbiologically positive or negative. Results: There were 1970 images from 886 patients were included. None of the models were better than random chance at predicting positive microbiologic results (area under the receiver operating characteristic curve ranged from 0.49 to 0.56; all confidence intervals included 0.5). Conclusions: These two state-of-the-art deep convolutional neural network architectures could not reliably predict whether a corneal ulcer would be microbiologically positive or negative based on clinical photographs. This absence of detectable morphological differences informs the future development of computer vision models trained to predict the causative agent in infectious keratitis using corneal photography. Translational Relevance: These deep learning models were not able to identify morphological differences between microbiologically positive and negative corneal ulcers. This finding suggests that similar artificial intelligence models trained to identify the causative pathogen using only microbiologically positive cases may have potential to generalize well, including to cases with falsely negative microbiologic testing.
Assuntos
Inteligência Artificial , Ceratite , Humanos , Estudos Transversais , Ceratite/diagnóstico , Redes Neurais de Computação , ÚlceraRESUMO
BACKGROUND: Seasonal outbreaks of infectious conjunctivitis remain a public health issue. Determination of outbreak etiologies in the context of a worldwide pandemic may provide useful information to guide public health strategies. The aim of this study was to identify pathogens associated with outpatient infectious conjunctivitis during the COVID-19 Delta surge. METHODS: This prospective study was conducted from April 2021 to September 2021. All outpatients presenting to the Aravind Eye Center (Madurai, India) with signs and symptoms consistent with acute infectious conjunctivitis were eligible. Three swabs were obtained from each participant: one from each conjunctiva and one from the anterior nares. Samples were processed for metagenomic RNA deep sequencing (RNA-seq). RESULTS: Samples from 106 study participants were sequenced. The most common presenting symptoms were tearing (86%) and itching (71%). Preauricular lymphadenopathy was present in 38% of participants. 20% of participants had close contacts with similar symptoms. Systemic symptoms such as coughing, runny nose, vomiting or diarrhea were uncommonly reported. 60% of all participants used some medicated eye drops upon enrollment. 75% of study participants demonstrated infection with human adenovirus D (HAdV-D). 11% of conjunctivitis was associated with SARS-CoV-2. 15% had no definitive pathogen detected. 8% of all participants had codetection of more than one pathogen on RNA-seq. CONCLUSIONS: During the COVID-19 Delta surge in India, HAdV-D was the most common pathogen associated with infectious conjunctivitis. SARS-CoV-2 was the second most common associated pathogen. Seasonal surveillance may be necessary for the determination of emerging and reemerging pathogens responsible for infectious conjunctivitis.
Assuntos
Adenovírus Humanos , COVID-19 , Conjuntivite , Humanos , SARS-CoV-2 , Estudos Prospectivos , Índia/epidemiologia , Conjuntivite/epidemiologia , Adenovírus Humanos/genética , Doença Aguda , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
BACKGROUND: Viral conjunctivitis (pink eye) can be highly contagious and is of public health importance. There remains significant debate whether SARS-CoV-2 can present as a primary conjunctivitis. The aim of this study was to identify pathogens associated with outpatient infectious conjunctivitis during the COVID-19 Delta surge. METHODS: This prospective study was conducted in the spring and summer months of 2021. 106 patients with acute conjunctivitis who presented to the Aravind Eye Center in Madurai, India were included. One anterior nasal swab and one conjunctival swab of each eye were obtained for each enrolled patient. Samples were subsequently processed for unbiased metagenomic RNA deep sequencing (RNA-seq). Outcomes included clinical findings and codetection of other pathogens with SARS-CoV-2 in patients with conjunctivitis. RESULTS: Among the 13 patients identified with human coronavirus RNA fragments in their swabs, 6 patients had SARS-CoV-2 infection, 5 patients had coinfections of SARS-CoV-2 and human adenovirus (HAdV), 1 patient had a coinfection with human coronavirus OC43 and HAdV, and 1 patient had a coinfection of Vittaforma corneae and SARS-CoV-2. 30% had bilateral disease and symptoms on presentation. Petechial hemorrhage was noted in 33% of patients with SARS-CoV-2 infection. No patients with SARS-CoV-2 or SARS-CoV-2 and HAdV infections had subepithelial infiltrates on presentation. All patients denied systemic symptoms. CONCLUSIONS: Among the patients presented with conjunctivitis associated with human coronavirus infection, over 50% of the patients had co-infections with other circulating pathogens, suggesting the public-health importance of broad pathogen testing and surveillance in the outpatient conjunctivitis population.
Assuntos
COVID-19 , Coinfecção , Conjuntivite , Humanos , SARS-CoV-2 , Coinfecção/epidemiologia , Pacientes Ambulatoriais , Estudos Prospectivos , Índia/epidemiologia , RNARESUMO
PURPOSE: The objective of this study was to compare the clinical and microbiological profiles of culture-proven pure Corynebacterium keratitis with mixed infection and their antibiotic susceptibility patterns over a 2-year period. METHODS: A retrospective analysis of culture-proven cases of Corynebacterium keratitis over a 2-year period was performed in 3 different tertiary eye care centers. All isolates were tested for antibiotic susceptibility in vitro using the disc-diffusion method for 7 antibiotics. RESULTS: Altogether 108 cases were identified as culture-positive Corynebacterium keratitis in 3 tertiary eye care centers. Of these, 60.2% (n = 65) and 39.8% (n = 43) of cases were due to pure Corynebacterium and mixed infection, respectively. The mean duration of symptoms was 23.2 ± 29.6 days. In the mixed-infection group, fungus was identified as the coexistent pathogen in 22 cases (51.1%). Ocular surface disorder was the most common risk factor (33.9%) in Corynebacterium keratitis. The most frequently isolated species was Corynebacterium amycolatum (22.2%) in both groups. Therapeutic keratoplasty was performed in 8.3% of cases. There was no significant difference in the outcome between the 2 groups. Cefazolin resistance was seen in 13.9% of patients, and all isolates were susceptible to vancomycin. The resistance pattern showed emerging resistance toward fluoroquinolone because the isolates were resistant to gatifloxacin (58.3%), moxifloxacin (47.2%), ciprofloxacin (54.6%), and ofloxacin (45.4%). CONCLUSIONS: Ocular surface disorder is the most common risk factor in Corynebacterium keratitis. Although fluoroquinolones are commonly used as first-line therapy in microbial keratitis, the in vitro resistance pattern indicates that these are less likely to be effective in infection with Corynebacterium species.
Assuntos
Coinfecção , Infecções Oculares Bacterianas , Ceratite , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Cefazolina , Ciprofloxacina/uso terapêutico , Coinfecção/tratamento farmacológico , Corynebacterium , Infecções Oculares Bacterianas/tratamento farmacológico , Infecções Oculares Bacterianas/microbiologia , Fluoroquinolonas/farmacologia , Fluoroquinolonas/uso terapêutico , Gatifloxacina , Humanos , Ceratite/tratamento farmacológico , Ceratite/microbiologia , Testes de Sensibilidade Microbiana , Moxifloxacina/uso terapêutico , Ofloxacino/uso terapêutico , Estudos Retrospectivos , Centros de Atenção Terciária , Vancomicina/uso terapêuticoRESUMO
PURPOSE: To evaluate the clinical and microbiological features of a large cohort with culture-confirmed fungal endophthalmitis across India. DESIGN: Cross-sectional, hospital-based, retrospective medical record review. PARTICIPANTS: Seven large tertiary eye care centers from different regions of India. METHODS: Patient data were pooled from electronic or physical medical records of each participating center. Fellowship-trained vitreoretinal specialists clinically managed all patients, and in-house microbiology laboratories performed all microbiological workups. The clinical and microbiological procedures were broadly uniform across all participating centers. The essential treatment consisted of vitreous surgery as well as intravitreal and systemic therapies with antifungal agents. MAIN OUTCOME MEASURES: Clinical outcome of the causative event and causative fungus. RESULTS: In the period from 2005 to 2020, 7 centers treated 3830 cases of culture-proven endophthalmitis, and of these, 19.1% (n = 730) were cases of culture-confirmed fungal endophthalmitis. It included 46.9% cases of postoperative (87.4% postcataract surgery), 35.6% of traumatic, and 17.5% of endogenous endophthalmitis. The fungi included 39.0% of Aspergillus (high prevalence in central, east, and south zones), 15.1% of Candida (high prevalence in west zone), and 15.9% of Fusarium (high prevalence in north and west zones). The time to symptom development was between 1 week and 4 weeks in more than one third of the patients, except in patients with traumatic endophthalmitis. Less than half of the patients had hypopyon on presentation. The presenting visual acuity (PVA) in most patients was <20/400. Nearly all patients needed vitrectomy and an average of 2 intravitreal injections of antifungal agents. At least 10% of eyes needed therapeutic keratoplasty, and up to 7% of eyes were eviscerated. After treatment, the final (best corrected) visual acuity (FVA) was >20/400 in 30.5% (n = 222) of eyes and >20/40 in 7.9% (n = 58) of eyes, and 12% (n = 88) of eyes lost light perception. A post hoc analysis showed the male sex to be significantly more associated with traumatic endophthalmitis than with postoperative (P < 0.0001) and endogenous (P = 0.001) endophthalmitis, more isolation of Candida species in patients with endogenous endophthalmitis than in those with postoperative (P = 0.004) and traumatic (P < 0.0001) endophthalmitis, better PVA in eyes with Candida species infection (P < 0.0001), and poorer FVA in eyes with Aspergillus species infection. CONCLUSIONS: Fungal endophthalmitis is not uncommon in India. The inclusion of antifungal agents with antibiotics as the first empirical intravitreal therapy before microbiological confirmation should be considered when a fungal infection is suspected.
Assuntos
Candidíase , Endoftalmite , Infecções Oculares Fúngicas , Antifúngicos/uso terapêutico , Candida , Candidíase/diagnóstico , Candidíase/tratamento farmacológico , Candidíase/epidemiologia , Estudos Transversais , Endoftalmite/diagnóstico , Endoftalmite/epidemiologia , Endoftalmite/terapia , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/epidemiologia , Fungos , Humanos , Masculino , Estudos RetrospectivosRESUMO
PURPOSE: The purpose of this article was to evaluate the impact of sample collection order on the diagnostic yield of metagenomic deep sequencing (MDS) for determining the causative pathogen in infectious keratitis. METHODS: We performed a cross-sectional diagnostic test evaluation among subjects with infectious keratitis at Aravind Eye Hospital in Madurai, India. All subjects underwent corneal scrapings of the affected eye to obtain potassium hydroxide smear, Gram stain, bacterial culture, and fungal culture, in this order. The order of MDS specimen collection relative to smear and culture samples was randomized and served as the primary predictor. Outcomes included the normalized copy number of pathogenic RNA detected by MDS, the proportion of MDS samples that were diagnostic, and the agreement of MDS results with cultures. RESULTS: MDS samples from 46 subjects with corneal ulcers were evaluated. MDS was positive in 33 subjects (76%) and had 74% overall agreement with culture results. There was no association between order of MDS sample collection and normalized copy number of genetic material detected (P = 0.62) or the likelihood of MDS positivity (P = 0.46). However, the likelihood of agreement between MDS and cultures decreased when MDS corneal swabs were collected after other diagnostic corneal scrapings (P = 0.05). CONCLUSIONS: The overall yield of MDS for detecting the cause of infectious keratitis was not affected by sample collection order. However, diagnostic agreement between MDS and cultures decreased when MDS samples were collected after other specimens. Additional investigation is warranted to determine whether this represents increased sensitivity of MDS compared with cultures or higher susceptibility to contaminants.
Assuntos
Córnea/microbiologia , DNA Bacteriano/análise , DNA Fúngico/genética , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Fúngicas/microbiologia , Ceratite/microbiologia , Metagenoma/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Bactérias/genética , Bactérias/isolamento & purificação , Estudos Transversais , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/genética , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/genética , Feminino , Fungos/genética , Fungos/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Ceratite/diagnóstico , Ceratite/genética , Masculino , Pessoa de Meia-Idade , Manejo de Espécimes/métodos , Adulto JovemRESUMO
PURPOSE: The purpose of this study is to determine the epidemiology, risk factors, clinical features, and treatment outcome of molecularly diagnosed Periconia keratitis. METHODS: Clinical records of all culture proven fungal ulcers with molecular identification suggestive of Periconia species who presented to a single tertiary referral center from January 2012 to December 2013 were retrospectively analysed. RESULTS: Among 1356 cases of keratomycosis, 8 (0.6%) patients were affected due to Periconia species. The mean age of presentation was 59 years with males (nâ¯=â¯6; 75%) were more commonly affected than females (nâ¯=â¯2; 25%). Significant history of trauma was present only in one patient. The infiltrate size was less than 5â¯mm in majority of patients 75% (nâ¯=â¯6). 50% (nâ¯=â¯4) responded to antifungal, 12.5% (nâ¯=â¯1) responded to antibacterial, 12.5% (nâ¯=â¯1) required therapeutic penetrating keratoplasty, 25% (nâ¯=â¯2) lost to follow up after first visit. The mean duration of treatment in healed cases was 20 days. CONCLUSION: This is the first report on Periconia sp causing human corneal ulcer. This study signifies the importance of molecular identification in the diagnosis of rare fungi which will improve our understanding on disease pathology and outcome. Visual prognosis appears good if the infection is diagnosed and topical antifungal interventions started early.
Assuntos
Ascomicetos , Úlcera da Córnea , Infecções Oculares Fúngicas , Micoses , Antifúngicos/uso terapêutico , Ascomicetos/patogenicidade , Úlcera da Córnea/tratamento farmacológico , Úlcera da Córnea/epidemiologia , Úlcera da Córnea/microbiologia , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/tratamento farmacológico , Infecções Oculares Fúngicas/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Micoses/tratamento farmacológico , Micoses/epidemiologia , Estudos RetrospectivosRESUMO
PURPOSE: To describe demographics, risk factors, antibiotic susceptibility, management and outcomes of ocular infections caused by non-tuberculous mycobacteria (NTM). METHODS: A retrospective review of medical case records and microbiology records of patients with ocular infections that were culture positive for non-tuberculous Mycobacteria from January 2014 to December 2018 was done. Antibiotic susceptibility profile was done based on the CLSI guidelines. Laboratory diagnosis for the NTM Species was done by conventional microbiological methods. The species identification was done for stored isolated utilizing polymerase chain reaction targeting 16S rDNA and rpoB gene, followed by DNA sequencing and phylogenetic analysis. RESULTS: Twenty patients with NTM ocular infections were identified during the study period. A majority of cases presented as 12 infectious keratitis (60%) and three suture-related corneal infiltrates (15%). Common risk factors were history of trauma in 9 (45%) patients and history of ocular surgery in 5 (25%) patients. Patients were treated with combination of amikacin and flouroquinolones/chloramphenicol (70%) and surgical interventions were performed in 25% cases. Only twelve isolates were stored and ten isolates were identified as the M. abscessus subsp. abscessus and two isolates as M. abscessus subsp. massiliense by sequencing and phylogenetic analysis. Majority of the NTM were sensitive to amikacin (75%) followed by moxifloxacin, ciprofloxacin, cephotaxime and tobramycin (35%). CONCLUSION: High degree of clinical suspicion, multidrug antibiotic therapy and timely surgical intervention in patients with NTM infections, are advised for better clinical outcomes. Prior ocular trauma, prior ocular surgery and presence of biomaterials were the major predisposing factors. Earlier surgical intervention in cases where abscesses or biomaterials are involved, is necessary for rapid recovery.
Assuntos
Infecções Oculares , Infecções por Mycobacterium não Tuberculosas , Amicacina/uso terapêutico , Antibacterianos/uso terapêutico , Materiais Biocompatíveis , Infecções Oculares/tratamento farmacológico , Infecções Oculares/epidemiologia , Infecções Oculares/microbiologia , Humanos , Índia/epidemiologia , Testes de Sensibilidade Microbiana , Infecções por Mycobacterium não Tuberculosas/diagnóstico , Infecções por Mycobacterium não Tuberculosas/tratamento farmacológico , Infecções por Mycobacterium não Tuberculosas/epidemiologia , Micobactérias não Tuberculosas/genética , Filogenia , Estudos RetrospectivosAssuntos
Úlcera da Córnea/diagnóstico , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Fúngicas/diagnóstico , Metagenômica , Infecções Estafilocócicas/diagnóstico , Infecções Estreptocócicas/diagnóstico , Úlcera da Córnea/microbiologia , DNA Bacteriano/genética , Testes Diagnósticos de Rotina , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Fúngicas/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Metagenoma , RNA Bacteriano/genética , Infecções Estafilocócicas/microbiologia , Staphylococcus/genética , Staphylococcus/isolamento & purificação , Infecções Estreptocócicas/microbiologia , Streptococcus/genética , Streptococcus/isolamento & purificaçãoRESUMO
PURPOSE: To explore the use of optical SmartProbes for the rapid evaluation of corneal scrapes from patients with suspected microbial keratitis, as a clinical alternative to Gram stain. DESIGN: Experimental study with evaluation of a diagnostic technology. METHODS: Corneal scrapes were collected from 267 patients presenting with microbial keratitis at a referral cornea clinic in South India. Corneal scrapes were flooded with SmartProbes (BAC One or BAC Two) and evaluated by fluorescence microscopy (without the need for sample washing or further processing). The SmartProbe-labeled samples were scored as bacteria/fungi/none (BAC One) or gram-negative bacteria/none (BAC Two) and compared to Gram stain results. RESULTS: Compared to Gram stain, BAC One demonstrated sensitivity and specificity of 80.0% and 87.5%, respectively, positive and negative predictive values (PPV, NPV) of 93.8% and 65.1%, and an accuracy of 82.2. BAC Two demonstrated sensitivity and specificity of 93.3% and 84.8%, respectively, an NPV of 99.2%, and an accuracy of 85.6%. When the corresponding culture results were compared to the Gram stain result, the sensitivity and specificity were 73.4% and 70.7%, the PPV and NPVs were 86.5% and 51.0%, and overall accuracy was 72.6. CONCLUSIONS: Fluorescent SmartProbes offer a comparative method to Gram stain for delineating gram-positive or gram-negative bacteria or fungi within corneal scrapes. We demonstrate equivalent or higher sensitivity, specificity, PPV and NPVs, and accuracy than culture to Gram stain. Our approach has scope for point-of-care clinical application to aid in the diagnosis of microbial keratitis.
Assuntos
Bactérias/isolamento & purificação , Córnea/microbiologia , Úlcera da Córnea/microbiologia , Infecções Oculares Bacterianas/microbiologia , Infecções Oculares Fúngicas/microbiologia , Corantes Fluorescentes , Fungos/isolamento & purificação , Corantes/química , Violeta Genciana , Humanos , Testes de Sensibilidade Microbiana , Microscopia de Fluorescência , Oxidiazóis/química , Fenazinas , Estudos Retrospectivos , Sensibilidade e EspecificidadeRESUMO
Introduction. Nocardia spp. can cause several ocular infections, such as keratitis, endophthalmitis and scleral abscesses. Molecular identification of Nocardia spp. by 16S rDNA sequencing is the gold standard method at present for species identification, but closely related species can only be identified by multilocus sequence analysis (MLSA) of housekeeping genes.Aim. The major objective was to profile Nocardia species, antibiotic susceptibility patterns and clinical outcomes in endophthalmitis patients.Methodology. Between January 2009 and December 2018, endophthalmitis patients who were diagnosed with Nocardia infection based on microscopic and culture characteristics were selected. Antibacterial susceptibility tests were performed and Nocardia speciation was performed using MLSA and phylogenetic tree analysis of the 16 s rRNA gene and the gyrB, hsp65 and secA1 genes.Results. A total of 43 culture-proven patients were identified during the study period. All isolates were 100â% sensitive to amikacin and 98â% resistant to ceftazidime. Fluoroquinolone sensitivity was observed in the range of 58 to 72â%. Year-wise analysis of antibiotic resistance patterns revealed there was a significant increase in resistance to fluoroquinolones. Twenty-two isolates were stored and six different species were identified. Nocardia farcinica (n=10) was found to be the most predominant, followed by Nocardia cyriacigeorgica (n=4), Nocardia otitidiscaviarum (n=3), Nocardia amikacinitolerans (n=2), Nocardia puris (n=2) and Nocardia higoensis (n=1).Conclusions. N. farcinica is the major pathogen, and this is the first report to identify N. otitidiscaviarum, N. amikacinitolerans and N. higoensis as causing endophthalmitis. Overall, visual outcomes were mostly poor even after aggressive management.
Assuntos
Endoftalmite/diagnóstico , Endoftalmite/microbiologia , Tipagem de Sequências Multilocus , Nocardiose/diagnóstico , Nocardiose/microbiologia , Nocardia/classificação , Nocardia/genética , Antibacterianos/farmacologia , Resistencia a Medicamentos Antineoplásicos , Endoftalmite/tratamento farmacológico , Genes Bacterianos , Genes Essenciais , Humanos , Nocardia/efeitos dos fármacos , Nocardiose/tratamento farmacológico , Filogenia , RNA Ribossômico 16S/genéticaRESUMO
Introduction. Acanthamoeba keratitis is a sight-threatening corneal infection that is commonly reported among contact lens users and those suffering from corneal trauma. The prevalence of Acanthamoeba species or genotypes in causing keratitis infection is not well known.Aim. This study was conducted to identify and genotype Acanthamoeba isolates from keratitis patients, targeting the ribosomal nuclear subunit (Rns) region, and describe the associated clinical presentation and treatment outcome.Methodology. Thirty culture-confirmed patients with Acanthamoeba keratitis, identified in a tertiary eye care centre in South India during the period from December 2016 to December 2018, were included in this study. The data collected from patient records include demographic details, history of illness, mode of trauma, treatment history and follow-up status. The genotype and species were identified based on the Rns sequence and phylogenetic tree analysis.Results. Acanthamoeba culbertsoni was the most predominant keratitis-causing species, followed by Acanthamoeba quina, Acanthamoeba castellanii, Acanthamoeba healyi, Acanthamoeba hatchetti, Acanthamoeba polyphaga and Acanthamoeba stevensoni. Three major genotypes were identified (T4, T11 and T12), with the T4 genotype being the most predominant, with four subclusters, i.e. T4A, T4B, T4D and T4E. This is the first report on corneal infection by the A. stevensoni T11 genotype and the A. healyi T12 genotype. No significant correlation was observed between the clinical outcomes of corneal disease and the genotypes or species.Conclusion. Rns genotyping is very effective in identifying the Acanthamoeba species and genotype in keratitis. Genotyping of Acanthamoeba spp. will help to advance our understanding of genotype-specific pathogenesis and geographical distribution.