Microsporidial keratitis: Literature review and report of 2 cases in a tertiary eye care center.

Microsporidia are getting more recognized as causative organism of ocular, gastrointestinal, renal, pulmonary, and sinus diseases, in both immunocompetent and immunosuppressed patients. Ocular microsporidiosis can be isolated or may present as part of systemic infection. Recent reports showed increasing number of cases of ocular microsporidiosis in immunocompetent individuals. The ocular type occurs mainly in two forms: keratoconjunctivitis form which is mostly seen in immunocompromised individuals and stromal keratitis form seen in immunocompetent individuals. The ocular cases which present with superficial keratitis in acquired immune deficiency syndrome (AIDS) patients differ from the cases seen in immunocompetent individuals which present mainly as deep stromal keratitis. We are presenting the only two documented cases of microsporidial keratitis diagnosed over 25 years of practice in our institution. The cases are diagnosed by identification of the Acid-fast organisms. Ultrastructure and additional information on species identification in one of the cases. Both cases are eventually managed by therapeutic PKP. Diagnosis and treatment modalities are discussed based on a comprehensive literature review.

Incidence and microbiological profile of mycotic keratitis in a tertiary care eye hospital: A retrospective analysis.

PURPOSE: To determine the incidence and microbiological profile of mycotic keratitis seen at a tertiary care eye hospital. MATERIALS AND METHODS: A retrospective review of microbiology records of patients presenting with suspected microbial keratitis seen between January 2006 and December 2009 was performed. Patients with positive fungal cultures were further analyzed for the type of fungus isolated and associated bacterial pathogens. RESULTS: Microbiology records of 2300 patients with suspected microbial keratitis were reviewed. A microbiological diagnosis of mycotic keratitis was established in 87 (3.8%) patients over a four year period based on positive fungal cultures. The yearly incidence of mycotic keratitis was 3.2% (2006), 4.9% (2007), 3.3% (2008) and 3.6% (2009). Filamentous fungi were isolated more often than yeasts. Aspergillus species followed by Fusarium species and Trichophyton species were the commonest filamentous fungi isolated while Candida albicans was the most frequently encountered yeast. Mixed infections due to fungal and bacterial pathogens were seen in 25/87 (28.7%) patients. CONCLUSION: Cumulative incidence of mycotic keratitis was 3.8% over a four year period. Aspergillus species and Candida albicans were the most frequent pathogenic organisms causing mycotic keratitis in this part of the world. Mixed infections were seen in 28.7% of the patients. Knowledge of the "local" etiology within a region may be valuable in the management of mycotic keratitis in instituting an empirical therapy, especially when facilities for microscopy, cultures and antifungal susceptibility are not readily available. The baseline information presented will also be helpful in the planning of a corneal ulcer management strategy and for future studies on mycotic keratitis.

Severe hemorrhagic retinopathy as initial manifestation of acute retinal necrosis caused by herpes simplex virus.

A case of bilateral acute retinal necrosis due to herpes simplex virus 1, in a child is reported. The case presented as an extensive hemorrhagic retinopathy that was misdiagnosed as non-infective initially. Diagnostic aqueous tap of the blind eye for viral DNA by polymerase chain reaction helped to confirm viral etiology when the other eye was affected. Appropriate antiviral therapy followed by prompt surgeries for subsequent retinal detachment helped to salvage useful vision in the second eye.

Relationship between clinical presentation and visual outcome in postoperative and posttraumatic endophthalmitis in south central India.

PURPOSE: To determine risk factors for poor visual outcome in postoperative and posttraumatic endophthalmitis in a large referral center in south central India. METHODS: In this prospective observational series the authors examined 388 patients of postoperative (n= 206) and posttraumatic (n= 182) endophthalmitis at the L V Prasad Eye Institute in Hyderabad, India between 1991 and 1997. The analysis was confined to 236 patients-128 (62.1%) postoperative and 108 (59.3%) posttraumatic patients who were followed for a minimum period of 3 months. A detailed protocol was followed. Chi-square and logistic regression analysis were used to determine risk factors for visual outcome worse than 6/18 and worse than 6/120. RESULTS: Postoperative endophthalmitis: In univariate analysis the features associated with poor visual acuity (grouped as < 6/18 and < 6/120) included intracapsular cataract surgery, poor presenting visual acuity, presence of vitreous cells, inability to visualise the optic disc on indirect ophthalmoscopy, presence of vitreous membranes on ultrasonography, and a culture-positive vitreous biopsy. In the multivariate analysis, visual acuity of less or equal light perception (LP) at presentation was associated with a 3-month postoperative visual acuity of < 6/18, with an odds ratio of 5.85 [1.25 - 27.42, 95% CI], and vitreous membranes seen on ultrasonography was associated with a final visual acuity of < 6/120, with an odds ratio of 2.47 [1.05 - 5.83, 95% CI]. Posttraumatic endophthalmitis: In univariate analysis the features associated with poor visual acuity (grouped as < 6/18 and < 6/120) included a retained intraocular foreign body (IOFB), trauma by needle (hypodermic or sewing), poor presenting visual acuity, inability to visualise the optic disc on indirect ophthalmoscopy, presence of vitreous membranes on ultrasonography, and a culture-positive vitreous biopsy. In multivariate analysis, IOFB was associated with a 3-month follow-up visual acuity of < 6/18, with an odds ratio of 5.90 [1.85 - 18.78, 95% CI], and trauma by a needle (hypodermic or sewing) and retained IOFB was associated with a final visual acuity of < 6/120, with an odds ratio of 4.47 [1.22 - 16.38, 95%CI] and 3.76 [1.36 - 10.37, 95% CI] respectively. CONCLUSION: This is the largest, single-centre, prospective study on risk factors for poor visual outcome in postoperative and posttraumatic endophthalmitis. The independent risk factor for 3-month follow-up visual acuity of < 6/18 was the presenting visual acuity of < or =LP in postoperative endophthalmitis and a retained IOFB in posttraumatic endophthalmitis. The independent risk factor for 3-month visual acuity of < 6/120 was the presence of vitreous membranes on ultrasonography in postoperative endophthalmitis, and trauma by a needle (hypodermic/ sewing) and retained IOFB in posttraumatic endophthalmitis.

Ocular and neuronal cell apoptosis during HSV-1 infection: a review.

HSV-1 may activate or suppress the apoptotic pathway in various cells. This review will discuss this apparent dichotomy and place particular emphasis on the different strategies HSV-1 uses to block or suppress the apoptotic pathway in various cell lines and tissues.

Multiplex polymerase chain reaction for the detection of herpes simplex virus, varicella-zoster virus and cytomegalovirus in ocular specimens.

PURPOSE: A majority of ocular viral diseases are caused by herpes group of viruses. Such infections, especially atypical herpetic keratitis, iridocyclitis and intra-ocular inflammations, can often present with overlapping clinical manifestations misleading the diagnosis. Molecular techniques are most useful in such instances for an accurate and rapid diagnosis since conventional methods are time consuming and less sensitive. A multiplex PCR was developed and used for the detection of herpes simplex virus (HSV), varicella zoster virus (VZV), and cytomegalovirus (CMV) in ocular samples. METHODS: One hundred and forty six ocular samples (corneal scrapings - 52, aqueous fluid - 36, vitreous fluid - 31, tissues - 26, skin vesicle scraping - 1) were included in the study. The sensitivity of the assay was determined using serial dilutions of standard strains of HSV, VZV, and CMV vis-à-vis plaque forming assay. RESULTS: The sensitivity of the assay was 4, 4 and 12 PFU/ml or 20, 20 and 60 genome copy numbers of HSV, VZV and CMV respectively. Using DNA from various sources (fungal, bacterial, human leukocytes, tissues) along with standard positive controls, the assay was found to be highly specific. HSV DNA was detected in majority of the clinical samples (33.6%), most frequent being corneal samples. Comparatively, VZV and CMV infections were detected in small number of samples (VZV-3, CMV-2). CONCLUSIONS: We found the assay very useful in our set-up whenever a differential diagnosis of herpetic infections was suggested by the ophthalmologist. The multiplex PCR we have described here can be of greater value in clinics with larger number of patients suspected of having HSV, VZV or CMV infections.

Evaluation of corneal scraping smear examination methods in the diagnosis of bacterial and fungal keratitis: a survey of eight years of laboratory experience.

PURPOSE: To determine the sensitivity, specificity, and predictive values of Gram and potassium hydroxide with calcofluor white (KOH+CFW) stains in the diagnosis of early and advanced microbial keratitis, a retrospective analysis of comparative data from a prospectively collected database was done. METHODS: Patients with nonviral microbial keratitis seen at L.V. Prasad Eye Institute between February 1991 and December 1998 were included in the study. The type of bacteria seen on Gram stain was determined from 251 corneal scrapings from patients with early keratitis and 841 corneal scrapings from patients with advanced keratitis. The presence of fungi in corneal scrapings was determined by KOH+CFW stain of 114 and 363 scrapings from patients with early and advanced keratitis, respectively. The smear findings were compared with culture results to analyze specificity, sensitivity, and predictive values of the staining techniques. RESULTS: The sensitivity of Gram stain in the detection of bacteria was 36.0% in early and 40.9% in advanced keratitis cases; however, the specificity was higher in both groups (84.9% and 87.1%, respectively). Comparatively, the sensitivity and specificity of fungal detection were higher using KOH+CFW in early (61.1% and 99.0%, respectively) as well as advanced keratitis (87.7% and 83.7%, respectively). Predictive values were high for KOH+CFW in fungus detection, while they were poor for Gram stain in bacteria detection. In advanced keratitis cases, the false positives were higher in fungal detection (16.3%) than in bacterial detection (10.3%), while the false negatives were significantly higher in bacterial detection compared with fungal detection (59.1% versus 12.3%, p< 0.0001). In early keratitis, on the other hand, both false positives and false negatives for bacterial detection were significantly higher than fungal detection. CONCLUSIONS: Decisions can reliably be based on KOH+CFW stain of corneal scrapings for initiation of antifungal therapy in mycotic keratitis. The results of Gram stain, on the other hand, have limited value in therapeutic decisions for bacterial keratitis. Therefore, the search for a better modality for early and efficient diagnosis of bacterial keratitis needs to continue.

The epidemiological features and laboratory results of fungal keratitis: a 10-year review at a referral eye care center in South India.

PURPOSE: To report the epidemiological features and laboratory results of 1,352 cases of fungal keratitis diagnosed at the L.V. Prasad Eye Institute (LVPEI) in south India. METHODS: The medical and microbiology records of 1,352 culture proven cases (1,354 eyes) of fungal keratitis diagnosed at the LVPEI between January 1991 to December 2000 was retrospectively reviewed for demographic features, risk factors, seasonal variation, and laboratory findings. RESULTS: Males (962) were affected significantly more (p< 0.0001) than females (390). Of 1,352 patients, 853 (64.4%) were in the younger age group (16-49 years). Ocular trauma predisposed to infection in 736 (54.4%) of 1,354 eyes. There was a higher incidence of fungal keratitis during the monsoon and winter than summer. A fungal cause was established by smears of corneal scrapings in 1,277 (95.4%) eyes. The potassium hydroxide preparation (KOH), Calcofluor white (CFW), Gram-, and Giemsa-stained smears revealed fungus in 1,219 (91.0%), 1,224 (91.4%), 1,181 (88.2%), and 1,139 (85.1%) eyes, respectively. Fusarium(506, 37.2%) and Aspergillus species (417, 30.7%) predominated the hyaline fungal spectrum (1,133) and Curvularia species (39, 2.8%) were the highest among the dematiaceous isolates (218). CONCLUSIONS: To the best of our knowledge, this review presents the epidemiological features and laboratory results of the largest series of fungal keratitis ever reported in the literature. Keratomycosis is predominant in young adults with trauma as the major predisposing factor. With fungal keratitis being a major ophthalmologic problem in the tropical regions of the world, data available on the epidemiological features of a large series would greatly help medical practitioners at primary and secondary health care centers in the management of the disease. A simple KOH preparation of corneal scraping alone is highly beneficial in confirming the diagnosis.

Comparison of an immortalized human corneal epithelial cell line with Vero cells in the isolation of Herpes simplex virus-1 for the laboratory diagnosis of Herpes simplex keratitis.

BACKGROUND: Herpes simplex keratitis (HSK) is a sight threatening ocular infection often requiring a specific and prompt laboratory diagnosis. Isolation of Herpes simplex virus (HSV-1) in culture provides the most reliable and specific method and is considered as the "Gold Standard" in the laboratory diagnosis of HSK in spite of its low sensitivity. Using "cell lines of corneal origin" for virus isolation may be beneficial under such circumstances, since these cells have been shown to be excellent substrates for the growth of HSV-1 isolated from the cornea. We report a comparative study of a novel human corneal epithelial cell line (HCE) and the Vero cell line in the isolation of HSV-1 from corneal scrapings employing a shell vial assay. METHODS: Corneal scrapings were obtained from 17 patients with a clinical diagnosis of HSK. All the cases were confirmed by virological investigations (PCR and viral antigen detection positive, n = 15, PCR positive, n = 1, Viral antigen positive, n = 1). Scrapings obtained from 10 patients with infectious keratitis of non-viral origin were included as controls. All the scrapings were simultaneously inoculated into shell vials of HCE and Vero cells. Cultures were terminated at 24 h post-infection. Isolation of HSV-1 was confirmed using an indirect immunofluorescence/ immunoperoxidase assay. RESULTS: Virus could be isolated using both or either of the cell lines in 10/17 (58.82%) patients with HSK. HSV-1 was isolated from 10/17 (58.82%) and 4/17(23.52%) specimens in HCE and Vero cells, respectively (P = 0.036). None of the controls yielded HSV-1. While all the 10 (100%) strains were isolated in HCE, Vero yielded only 4/10 (40%) strains in the shell vial culture (P = 0.014). CONCLUSIONS: HCE showed a statistically significant difference in the virus isolation rate with respect to Vero cells. HCE may be an excellent alternative cell line for the isolation of HSV-1, especially from corneal scrapings, for the laboratory diagnosis of HSK.

Comparison of the sensitivity of a 24 h-shell vial assay, and conventional tube culture, in the isolation of Herpes simplex virus - 1 from corneal scrapings.

BACKGROUND: Herpes simplex keratitis is a sight threatening ocular infection. A rapid and specific diagnosis is essential for the institution of specific antiviral therapy and to avoid complications that can arise from misdiagnosis and inappropriate treatment. Though a variety of techniques are available, isolation of Herpes simplex virus 1 (HSV-1) in culture provides the most reliable and specific method, and is considered as the gold standard in laboratory diagnosis of herpes simplex keratitis. We report a comparative study of the sensitivity of a 24 h-shell vial assay and conventional tube culture in the isolation of HSV-1 from corneal scrapings. METHODS: A total of 74 corneal scrapings obtained from 74 patients with a clinical suspicion of herpes simplex keratitis submitted for the isolation of HSV-1, were simultaneously inoculated into shell vial and tube cultures employing the vero cell line. Shell vial and tube cultures were terminated at 24 h and fifth day respectively. Isolation of HSV-1 was confirmed employing an indirect immunofluorescence assay. RESULTS: HSV-1 was isolated from 24/74 (32.4%) specimens employing both the methods. Sensitivity of both the techniques were found to be similar (20/24, 83.3%) (P = 1.0). CONCLUSION: A 24 h-shell vial assay is a rapid alternative technique in comparison to the time consuming conventional tube cultures for the isolation of HSV-1, especially from corneal scrapings for the laboratory diagnosis of herpes simplex keratitis.