Nested polymerase chain reaction (nPCR) was performed on enucleated eyeball for detection of Mycobacterium tuberculosis (M. tb) genome in a patient with Eales' disease. PCR analysis in all previous studies has been done mainly using aqueous, vitreous and epiretinal membranes from these patients. Paraffin wax embedded tissue section of the enucleated eyeball was analyzed by histopathology and nPCR targeting MPB64 gene and IS6110 region of M. tb genome. Lymphocytic infiltration was seen in the vitreous, iris and the retinal tissue. Ziehl Neelsen stain was negative for acid fast bacilli. Caseation necrosis was not seen in any section. Agarose gel electrophoretogram showed positive results with 200 bp specific amplified product targeting MPB64 gene, whereas nPCR targeting IS6110 region was negative. Since biopsy proven M. tb is extremely difficult in ocular tissues due to extensive necrosis, the nPCR technique aided in the diagnosis.

Purpose: To report a case of Acanthamoeba endophthalmitis after an uneventful cataract surgery. Methods: Description, management, and outcomes of a biopsy-proven case of Acanthamoeba endophthalmitis. Results: Two days after a routine cataract surgery elsewhere, the patient presented with acute endophthalmitis diagnosed as a bacterial infection based on timing and severity. When conventional methods of management failed, the patient was referred to us. Only the presence of an atypical ring infiltrate suggested Acanthamoeba as a probable cause. Subsequent diagnostic evaluations confirmed the initial suspicion. Management with topical antiamoebics and intracameral and intravitreal voriconazole was attempted. Systemic voriconazole and metronidazole were also administered. However, because of relentless disease progression, the eye had to be eviscerated. The cornea, aqueous, vitreous, and sclera were positive by culture and/or polymerase chain reaction and histopathology. Conclusions: Acanthamoeba usually causes a chronic smoldering keratitis and, very rarely, scleritis. This report, which is the first of its kind, emphasizes the fact that fulminant endophthalmitis with associated scleritis can occur after ocular surgery in an immunocompetent individual, with no significant ophthalmic history.

All of the 100 clinical specimens from the control group were negative for both nPCRs. Out of the 354 clinical specimens, 339 were positive for both culture and nPCRs, 10 and 5 were positive for culture, and nPCR targeting IS6110 and MPB64 regions, respectively. To conclude, nPCRs targeting MPB64 and IS6110 region are reliable and specific targets when applied simultaneously on clinical specimens to attain 100% sensitivity for the detection of M. tuberculosis genome.