Microsporidial keratitis in India: 16S rRNA gene-based PCR assay for diagnosis and species identification of microsporidia in clinical samples

Purpose: To evaluate 16S rRNA-based polymerase chain reactions for the detection and species identification of the microsporidia that cause keratitis. Methods: Of the 5892 cases of microbial keratitis seen between September 2002 and December 2005, 31 (0.5%) microscopically diagnosed cases of microsporidial keratitis were included in the test group; 103 patients with nonmicrosporidial keratitis constituted the control group. A 16S rRNA-based pan-microsporidian PCR was chosen for the detection of microsporidian DNA. Species level identification was made using species-specific primer sets of Encephalitozoon spp (E. cuniculi, E. hellem, and E. intestinalis). Sequencing and BLAST analysis of amplicons obtained with pan-microsporidian primers were performed for validation. Results: The corneal scrapings from 26 of 31 cases in the test group and 2 of 103 cases in the control group showed a 250- to 280-bp amplicon in PCR by pan-microsporidian primers (sensitivity of 83% and specificity of 98%). The amplicons of 13 of 26 test group samples were identified by species-specific PCR: E. cuniculi, n = 7 (549 bp); E. hellem; n = 3 (549 bp); E. intestinalis; n = 1 (520 bp). The two cases in the control group were identified to be E. cuniculi. The remaining 15 cases (test group) were confirmed to be Vittaforma corneae by sequencing and BLAST analysis. All species were confirmed by sequencing and database homology comparison. Conclusions: This study is the first to validate PCR-based assays for detection of microsporidial DNA in corneal scrapings. Pan microsporidian PCR can be a useful adjunct with smear examination in the diagnosis of microsporidial keratitis

Update on Pathologic Diagnosis of Corneal Infections and Inflammations

One of the most frequent types of corneal specimen that we received in our pathology laboratory is an excised corneal tissue following keratoplasty. Several of these cases are due to corneal infections or the sequelae, like corneal scar. Advances in the histological and molecular diagnosis of corneal infections and inflammations have resulted in rapid and accurate diagnosis of the infectious agent and in the overall understanding of the mechanisms in inflammatory diseases of the cornea. This review provides an update of histopathological findings in various corneal infections and inflammations

Use of Different Stains for Microscopic Evaluation of Corneal Scrapings for Diagnosis of Microsporidial Keratitis

Retrospective evaluation of potassium hydroxide plus calcofluor white (KOH+CFW), Gram, Giemsa, and modified Ziehl-Neelsen (1% H(2)SO(4), cold) stains for the detection of microsporidia in corneal scrapings from 30 patients showed KOH+CFW and acid-fast stains to be most efficient (29/30 [96.7%] and 28/30 [93.3%], respectively) in the diagnosis of microsporidial keratitis

Calotropis-induced toxic keratitis managed with teleophthalmology services in rural India

A female laborer presented with acute diminution of vision in her right eye following accidental injury with a sap of Calotropis flower. The patient was diagnosed with Calotropis-mediated endothelial toxicity based on clinical findings and started on a course of topical steroids. The patient had to face several logistical challenges such as distance of travel, loss of daily wages, and nonavailability of a family member to accompany her. She was then followed up with sequential imaging of the cornea utilizing the teleophthalmology services of our institute at the rural vision center located in the vicinity of patient's house. Seven days later, the cornea had completely cleared with improvement in the best-corrected visual acuity. This case demonstrates Calotropis-induced endothelial toxicity which is very common in remote rural areas and its effective management using the mobile or tablet-based teleophthalmology application of our institute's three-tier eyecare pyramid model

Congenital anomalies of lens shape

The crystalline lens is an important structure in the eye that starts to develop as early as the 22nd day of gestation, with further differentiation that continues after the induction. Congenital anomalies of the lens may involve the size, shape, and position of the lens. They may sometimes be associated with anterior segment dysgenesis or persistence of the tunica vasculosa lentis and hyperplastic vitreous and hyaloid system. Manifestations of anomalies of the lens shape are usually seen in early or late childhood however may sometimes be delayed into adulthood based on the level of visual impairment or the presence or absence of any syndromic associations. While lens coloboma has more often been reported in isolation, the more commonly implicated genes include the PAX6 gene, lenticonus in particular anterior is often part of Alport syndrome with extra-ocular manifestations in the kidneys and hearing abnormalities due to mutations in the alpha 5 chain of the Type IV collagen gene. Recognition of these manifestations and obtaining a genetic diagnosis is an important step in the management. The level of visual impairment and amblyopia dictates the outcomes in patients managed either conservatively with optical correction as well as surgically where deemed necessary. This review discusses the various anomalies of the lens shape with its related genetics and the management involved in these conditions

Anterior megalophthalmos: Is visual restoration possible?

We report two cases of anterior megalophthalmos with cataract. Both cases have megalocornea, cavernous anterior chamber, enlarged iris–lens diaphragm, and normal axial length. The vision was less due to cataract. To restore vision, phacoemulsification was performed in each eye in both cases. Intraoperatively, to overcome anatomical challenges, we made scleral tunnel incision, stained anterior capsule, and fixated intraocular lens (IOL) by different techniques. In the first case, IOL was fixated through the sclera, whereas in the second case, IOL stabilization was achieved by capturing the optic in anterior capsulorhexis margin and placing the haptics in sulcus. Successful vision was restored in both cases without pseudophacodonesis

Corneal triple procedure: indications, complications, and outcomes: a developing country scenario

Purpose: We report the indications, complications, and outcomes of 104 corneal triple procedures in our institute. Methods: Patient records of 104 consecutive cases of corneal triple procedure (penetrating keratoplasty with extracapsular cataract extraction and intraocular lens implantation) performed by experienced corneal surgeons between January 1992 and December 1997 were retrospectively reviewed. Relevant preoperative, operative, and postoperative data were collected. The outcome was assessed by the graft clarity and visual acuity at the last visit. Survival analysis of these grafts was determined by Kaplan-Meier method. Results: Of 104 patients, 70 were men and 34 were women. Mean age of these patients was 48.5 ± 17.1 years (range, 1-75 years). Corneal scarring with cataract was the reason for surgery in 69 (66.4%) cases. The intraoperative complications included vitreous upthrust in seven (6.9%) cases and posterior capsular dehiscence in three (2.9%) cases. The most common early postoperative complications were increased intraocular pressure in 19 (18.3%) and increased anterior chamber reaction in 25 (24%) cases. Posterior capsular opacification was seen in 26 (25%), graft reaction in 15 (14.4%), and secondary glaucoma in 15 (14.4%) cases. These formed the important late postoperative complications. The average postoperative follow-up was 23.7 ± 17.6 months (range, 1.6-79.4 months); at which time 72% of the grafts remained clear. At last follow-up, 40% of patients had a visual acuity of ≥20/40. Conclusions: Corneal scarring with cataract is the most common reason for triple procedure in this part of the world. This is a safe surgical procedure with good graft clarity and reasonable visual recovery

Scleral thickness in normal Indian eyes measured using spectral domain anterior segment optical coherence tomography

Purpose: To establish normative data on anterior scleral thickness using the spectral domain anterior segment optical coherence tomography (AS-OCT). Methods: In total, 200 eyes of 100 healthy subjects underwent AS-OCT scans in the temporal and nasal quadrants. The scleral + conjunctival complex thickness (SCT) was measured by a single examiner. Mean SCT was analyzed for differences across age groups, gender, and location (nasal versus temporal). Results: Mean age was 46.4 ± 18.3 (21–84) years; male to female ratio was 54:46. Mean SCT (nasal + temporal) of the right eye (RE) was 682.3 ± 64.2 μm in males and 660.6 ± 57.1 μm in females. In the left eye (LE), it was 684.6 ± 64.9 μm in males and 661.8 ± 49.3 μm in females. These differences between male and female for both eyes were statistically significant (P = 0.006 and P = 0.002). The mean SCT of temporal and nasal quadrants in the RE was 678.54 ± 57.50 and 666 ± 66.2 μm, respectively. In the LE, the temporal mean SCT quadrant was 679.6 ± 55.8 μm, and the nasal was 668.6 ± 63.6 μm. Age had a negative correlation with SCT (−0.62 μm/year; P = 0.03), and males had a higher temporal SCT than females (22 μm higher; P = 0.03). After adjusting for age and gender in a multivariate analysis, temporal SCT was significantly (P < 0.001) higher than nasal SCT. Conclusion: In our study, mean SCT decreased with age and males had a higher temporal SCT. This is the first study to evaluate scleral thickness in the Indian population, and the data can be used as a baseline for comparing variations in scleral thickness in disease

Outcomes of keratoplasty in lattice corneal dystrophy in a large cohort of Indian eyes

Purpose: The purpose of this study is to evaluate the outcomes of keratoplasty for lattice corneal dystrophy (LCD) performed at a tertiary eye care center. Methods: A retrospective review of medical records of those patients who were clinically diagnosed to have LCD (72 eyes of 57 patients) and underwent either penetrating keratoplasty (PK, 58 eyes of 46 patients) or deep anterior lamellar keratoplasty (DALK, 14 eyes of 13 patients) between the years 1987 and 2014 was performed. The main outcome measures included demographics, clinical features, and outcomes of keratoplasty. Results: The median follow-up after keratoplasty was 3.1 years (interquartile range [IQR], 9 months to 9 years). The median best-corrected visual acuity (BCVA) was 0.18 (IQR, 0.10–0.48) (Snellen equivalent 20/30 [IQR, 20/25–20/60]) at 4 years postoperatively and 0.65 (IQR, 0.18-0.95) (Snellen equivalent 20/89 [IQR, 20/30–20/178]) at 10 years following surgery. DALK eyes had a significantly better BCVA than PK eyes at 2 years following keratoplasty. The median overall survival of grafts was 15.8 years. Late complications included recurrence of LCD (14 eyes), graft infiltrate (23 eyes), graft rejection (15 eyes), graft failure (16 eyes), and glaucoma (14 eyes). Conclusion: The outcomes of graft are similar following PK and deep anterior lamellar keratoplasty; however, the latter appears to provide slightly better visual outcome. Recurrence of dystrophy in the graft and graft infiltrates limit the overall graft survival in both the groups