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K.L. Therese, J. Therese, H.N. Madhavan; Development and application of a seminested Polymerase chain reaction for detection of Mycobacterium fortuitum and Mycobacterium chelonae on sections of epitretinal membrane (ERM) OF Eales disease . . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1676.
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Purpose:Eales disease is an idiopathic condition with a primary retinal perivasculitis of unknown aetiology, affecting the young adult Asian population in the age group of 15–40years. In our earlier studies the association of M. tuberculosis in significant number of vitreous aspirates 1,3. and epiretinal membrane (ERM)2. of Eales patients has been demonstrated . The present study was undertaken to determine the role of rapid growers of mycobacteria – Mycobacterium chelonae (M.chelonae) and Mycobacterium fortuitum (M.fortuitum) in pathogenesis of Eales disease. Methods: A seminested Polymerase chain reaction (snPCR) was developed by designing a new set of inner round primer sequence separately for M.fortuitum and M.chelonae and optimized following standard procedures. The standardized technique was applied on paraffin sections of 33 ERM ( 20 from Eales patients and 13 from non Eales patients as controls).In addition the nested PCR coding for the MPB 64 gene2 for detection of M.tuberculosis was also performed. Results: The snPCRs were specific to amplify M. fortuitum and M.chelonae, and sensitive to detect 1.5pg of M. fortuitum and 10 pg of M. chelonae DNA. Among the 20 Eales ERM specimens 14 ( M fortuitum –5, M chelonae–1 and both –1, M. tuberculosis–5,and both M. fortuitum and M. tuberculosis – 2)were positive for one or two of the three mycobacterial genomes. Among 13 control ERMs 4 (2 each of M.fortuitum and M.tuberculosis) were positive and other 9 were negative for all 3 mycobacteria. The difference between the two groups were statistically significant (P = 0.027.) Conclusions: The snPCR developed in this study is the first of its kind in literature for detection M.fortuitum and M. chelonae. Apart from M.tuberculosis, M. fortuitum and M. chelonae may also be associated with the pathogenesis of Eales disease. 1.Biswas J, Therese KL ,Madhavan HN . Br, J. of.Ophthalmol. 1999 ;83 994. 2.Madhavan HN,Therese KL,ET AL Invest.OpthalmolVis.Sci2000 41822–825 3.Madhavan HN, Therese KL Kavitha D Ind.J.of.Ophthalmol. 2002;50:35–39
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