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Savitri Sharma - One of the best experts on this subject based on the ideXlab platform.
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evaluation of multiplex real time polymerase chain reaction for the detection of herpes simplex virus 1 and 2 and varicella zoster virus in corneal cells from normal subjects and patients with Keratitis in india
Indian Journal of Ophthalmology, 2019Co-Authors: Sai Jeevana Madhuri Guda, Savitri Sharma, Bhavani Sontam, Bhupesh Bagga, Konduri Ranjith, Joveeta JosephAbstract:Purpose: To determine the presence of herpes simplex virus and varicella zoster virus (HSV 1 and 2, VZV) in the cornea of normal subjects by multiplex real time quantitative (qPCR) assay and evaluate its utility in the diagnosis of Viral Keratitis. Methods: Corneal epithelial cells from 33 eyes of 22 patients undergoing photorefractive keratectomy surgery (controls) and 50 corneal scrapings from 50 patients with suspected HSV Keratitis were analyzed for the presence of HSV1 by conventional PCR and for presence of HSV1 and 2 and/or VZV by multiplex real-time PCR. Corneal scrapings of patients were also tested for HSV1 antigen by immunofluorescence assay (IFA). The results were compared and clinical records reviewed. Results: HSV1 and VZV DNA were detected in 8/33 controls (mean-14.3 ± 7.96, range: 3-29.1 copies/mL) and 2/33 controls (mean-10.7 ± 10.9, range 3-18.5 copies/ml) respectively. HSV2 was not detected in any of the controls. Copy numbers above the mean + 1SD of controls were considered significant for Viral load in patient samples. Significantly higher number of corneal scrapings (39/50, 78%) from patients were positive for HSV1 (1.2 × 106 copies/mL ± 3.7 × 106 copies/mL) by real time qPCR compared to IFA (11/48, 23%, P value 0.0001) and conventional PCR (20/50, 40%, P value 0.0002). Double infection with HSV-1 (1.5 × 107 copies/ml) and HSV-2 (3.57 × 104 copies/ml) in one case and VZV infection (1.03 × 102 copies/ml) in another was also detected by the multiplex real-time PCR. Conclusion: Multiplex real-time PCR reliably detects HSV1 and 2 and VZV DNA and is ideal for the diagnosis of HSV and VZV Keratitis in an ocular microbiology laboratory.
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Reactivation of herpes simplex Viral Keratitis following the botulinum toxin injection
Wolters Kluwer Medknow Publications, 2018Co-Authors: Muralidhar Ramappa, Peter Y Jiya, Sunita Chaurasia, Milind Naik, Savitri SharmaAbstract:We describe a case of 55-year-old male farmer presented with recurrent corneal abrasions with a spastic entropion in the left eye. Superior cornea showed typical nummular opacities suggestive resolved herpetic eye diseases. On further enquiry, he had similar episodes in the past. Contralateral eye was essentially normal. Following the botulinum toxin injection for the management of spastic entropion, subject developed reactivation of herpetic necrotizing stromal Keratitis. Diagnostic corneal scrapings were negative for herpes simplex virus-1 antigen by immunofluorescence assay and for DNA by molecular techniques. The case was successfully managed with topical steroids and antiViral medications
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coexistence of herpes simplex virus infection in microsporidial stromal Keratitis associated with granulomatous inflammation
Indian Journal of Ophthalmology, 2017Co-Authors: Ruchi Mittal, Praveen Kumar Balne, Srikant K Sahu, Sujata Das, Savitri SharmaAbstract:Background: Microsporidial stromal Keratitis poses several diagnostic challenges. Patients may present with corneal ulceration, marked stromal thinning, or even as a quite corneal scar. The presentation of microsporidial stromal Keratitis commonly mimics Viral Keratitis. Microbiology scrapings are usually helpful; however, scraping and culture-negative cases pose a significant diagnostic dilemma. Histopathological examination is diagnostic but shows varying degree of inflammation, predominantly composed of polymorphonuclear leukocytes. Granulomatous inflammation, in microsporidial stromal Keratitis, is never well described, and the authors in this article aim to describe the presence of granulomatous inflammation in microsporidial stromal Keratitis, in patients with associated herpes simplex virus (HSV) Keratitis. Methods: This was a retrospective and observational study conducted at a tertiary eye care center. Results: Of 263 patients who underwent therapeutic penetrating keratoplasty for infectious Keratitis, during 2011–2013, seven patients were diagnosed as microsporidial stromal Keratitis. Microsporidial spores could be demonstrated on microbiological scrapings in 5/7 (71%) of cases, but identified on histopathological examination and also confirmed on polymerase chain reaction (PCR) for microsporidium in 100% of cases. There was evidence of diffuse stromal necrosis with markedly severe degree of polymorphonuclear leukocytic infiltrates, with granulomatous inflammation in 42% of cases. Interestingly, these were positive for HSV-1 DNA on PCR. Review of medical records revealed much severe clinical presentations in patients with granulomatous inflammation, in comparison to cases without granulomatous inflammation. Conclusions: The authors hereby recommend that severe clinical presentation in patients with microsporidial stromal Keratitis, markedly dense polymorphonuclear leukocytic infiltrates or the presence of granulomatous inflammation on the histopathological examination, should be investigated further for the presence of HSV-1 DNA for better patient management and good visual outcome.
Sheraz M Daya - One of the best experts on this subject based on the ideXlab platform.
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penetrating keratoplasty outcomes from a corneal unit compared to national data
British Journal of Ophthalmology, 2006Co-Authors: P Beckingsale, I Mavrikakis, N Alyousuf, E Mavrikakis, Sheraz M DayaAbstract:AIMS: To determine long term graft survival rates and visual results for different indications for penetrating keratoplasty from a single institution over 10 years and compare these to national outcome data. METHODS: Retrospective chart analysis. 784 records were available for review of 1096 consecutive penetrating keratoplasty procedures performed between 1990 and 1999 (72%). Outcomes of graft survival, visual acuity, and astigmatism were analysed and compared to national outcome data supplied by the UK Transplant Service. RESULTS: At 5 year follow up, overall graft survival was 66%. This was subdivided into 98% for keratoconus, 86% for Viral Keratitis, 85% for Fuchs' dystrophy, 84% for pseudophakic bullous keratopathy, 55% for regrafts, and 57% for other diagnoses. There was a significantly higher graft survival rate for all diagnostic subgroups except Fuchs' dystrophy at 3 years of follow up compared to the national average. Best corrected visual acuity at 5 years was 6/18 or better in 53% of cases. The mean keratometric astigmatism was 3.4 dioptres. CONCLUSION: Penetrating keratoplasty is a safe and effective treatment for selected corneal disorders. Penetrating keratoplasty for Viral Keratitis may achieve good results with long term antiViral treatment. Patients may achieve better outcomes if their surgery is performed at specialist centres.
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penetrating keratoplasty indications over a 10 year period
British Journal of Ophthalmology, 2004Co-Authors: N Alyousuf, I Mavrikakis, E Mavrikakis, Sheraz M DayaAbstract:Aims: To determine the indications for penetrating keratoplasty (PK) at the Corneoplastic Unit and Eye Bank, UK, a tertiary referral centre, over a 10 year period. Methods: Records of all patients who underwent PK at our institution between 1990 and 1999 were reviewed retrospectively. Of the 1096 procedures performed in this period, 784 records were available for evaluation (72%). Results: Regrafting was the most common indication, accounting for 40.9% of all cases. Keratoconus was the second most common indication (15%), followed by Fuchs’ endothelial dystrophy (9.3%), pseudophakic bullous keratopathy (7.6%), and Viral Keratitis (5.9%), which included both herpes simplex and herpes zoster and showed a statistically significant decreasing trend using regression analysis (p Conclusion: Regrafting is the leading indication for PK; Viral disease—although declining—is the leading primary diagnosis.
J F Lindo - One of the best experts on this subject based on the ideXlab platform.
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acanthamoeba infection as a cause of severe Keratitis in a soft contact lens wearer in jamaica
American Journal of Tropical Medicine and Hygiene, 2005Co-Authors: Zoe Wynterallison, Jacob Lorenzo Morales, Donovan Calder, Kraig Radlein, Antonio Ortegarivas, J F LindoAbstract:We report the case of a 29-year-old Jamaican patient who presented with severe pain, redness, and swelling of both eyes. She was a regular soft contact lens wearer who did not maintain standard lens care. She was treated for a possible microbial/Viral Keratitis using topical ciprofloxacin drops, topical acyclovir ointment, and topical atropine drops. The response was inadequate, and scrapings from her cornea, contact lens cases, and both lenses revealed Acanthamoeba on microscopy, which was shown to be Acanthamoeba polyphaga using polymerase chain reaction. She was treated using chlorhexidine 0.02% hourly, ciprofloxacin every 4 hours, and atropine 1% every 12 hours, along with oral ketoconazole 200 mg twice daily with a dramatic response. However, she subsequently suffered slow corneal epithelial regrowth with severe scarring, vascularization, and cortical lens opacification and was referred for penetrating keratoplasty and cataract surgery. This is the first case of severe Keratitis caused by Acanthamoeba to be reported from Jamaica and demonstrates that this emerging pathogen can be a cause of severe Keratitis in the tropics.
Zeng Yuanhong - One of the best experts on this subject based on the ideXlab platform.
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clinical observation and efficacy comparison of Viral Keratitis
Hainan Medical Journal, 2011Co-Authors: Zeng YuanhongAbstract:Objective To investigate the clinical features of Viral Keratitis and the efficacies of different treatment.Methods 54 cases with Viral Keratitis were randomly divided into treatment group(28 cases,30 eyes) and control group(26 cases) from March 2007 to May 2010,The control group was given oral antiViral medicine and eye treatment,and the treatment group was treated with traditional Chinese medicine and western medicine.Results The clinical curative rate and total effective in the treatment group were significantly higher than that in the control group,with P0.05;after the treatment,the recovery of visual acuity in the treatment group was better than that in the control group,with P0.05;the time of healing and curing of the treatment group were shorter than that of the control group,with P0.05.Conclusion Integrated traditional Chinese and western medicine to cure Viral Keratitis is more effective than the western medicine only,which is worth promoting.
Namrata Sharma - One of the best experts on this subject based on the ideXlab platform.
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quantitative analysis of herpes simplex virus 1 transcript in suspected Viral Keratitis corneal buttons and its clinical significance
Indian Journal of Ophthalmology, 2021Co-Authors: Shahzan Anjum, Seema Sen, Rinky Agarwal, Namrata Sharma, Seema Kashyap, Anjana SharmaAbstract:Purpose: The evaluation of Herpes Simplex virus-1 (HSV-1) transcript by different investigative methods (qPCR, PCR and IHC) in corneal buttons from suspected Viral Keratitis patients and the comparison of results with histopathological findings and clinical diagnosis. Methods: Sixty corneal buttons, 30 suspected Viral Keratitis, and 30 controls (keratoconus and bullous keratopathy) obtained after primary penetrating keratoplasty, were included in the study. All the corneal buttons were subjected to reverse transcriptase quantitative PCR (qPCR) for the detection of latency-associated transcript (LAT) gene, conventional PCR for polymerase (pol) gene, and immunohistochemistry (IHC) for HSV-1 antigen respectively. After obtaining baseline preoperative clinical data, all the patients were followed up for three years. The results obtained were correlated with clinicopathological features and follow-up data. Results: Of the 30 suspected Viral Keratitis patients there were 6 females and 24 males with mean age 46.5 ± 24.62 years (3-80 yrs). There was a marked male preponderance (80%). HSV-1 LAT transcript was detected in 23% (7/30) corneal buttons by qPCR, HSV-1 DNA in 6.7% (2/30) and HSV-1 antigen in 30% (9/30) cases by conventional PCR and IHC respectively. A statistically significant association was found between qPCR and DNA PCR (P = 0.04). All the 30 control corneas were negative for HSV-1 LAT gene, DNA and antigen. Conclusion: Detection of HSV-1 LAT transcript by qPCR may be superior to HSV-1 DNA PCR (conventional) and IHC, which has low sensitivity. However, the utility of HSV-1 LAT mRNA analysis as a diagnostic modality by qPCR needs to be validated on a larger patient cohort.
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evaluation of tear samples for herpes simplex virus 1 hsv detection in suspected cases of Viral Keratitis using pcr assay and conventional laboratory diagnostic tools
British Journal of Ophthalmology, 2011Co-Authors: Gita Satpathy, Anjana Sharma, Abhishek Kumar Mishra, Radhika Tandon, Manoj Sharma, Niranjan Nayak, Jeewan S Titiyal, Namrata SharmaAbstract:Background Herpes Simplex Virus (HSV) Keratitis is a leading cause of corneal blindness. Definitive laboratory diagnosis is essential for timely management. Collection of corneal scrapings in patients with advanced epithelial Keratitis and corneal thinning poses perforation risks; tear fluid is a feasible and convenient alternative but has not been widely evaluated for HSV detection. Methods Tear fluid alone (229) or along with corneal scrapings (153) from patients of suspected herpetic Keratitis was tested for HSV-1 antigen by indirect immunofluorescence assay, virus isolation in Hep 2 cells and PCR to amplify the 111 bp region of the thymidine kinase (tk) coding gene and the 144 bp region from the DNA polymerase coding gene of HSV. Results HSV 1 antigen was detected in 31/229 (13.53%) tear specimen and 35/153 (22.87%) corneal scrapings in immunofluorescence assay; virus was isolated from 12/229 (5.2%) tear and 17/153 (11.11%) corneal scrapings, and PCR was positive for both the genes in 32/229 (13.97%) tear specimen and 56/153 (36.66%) corneal scrapings. Conclusion Corneal scrapings yielded a significantly better HSV positivity than tears in both the PCR assay (p
