Development and application of double round multiplex polymerase chain reaction in the identification of microbial causes for prompt etiologic diagnosis of intra-vitreal aspirates from patients clinically diagnosed as endophthalmitis
Professor, Aravind Eye Hospital and Postgraduate Institute of Ophthalmology, Tamil Nadu
CO-Principal Investigator
Dr. S. Ramesh
Associate Professor and Head, Postgraduate Sri Paramakalyani College, Tamil Nadu, Dr. C. Shivkumar, Chief, Aravind Eye Hospital and Postgraduate Institute of Ophthalmology, Tamil Nadu, Dr. V. Maneksha, Chief, Aravind Eye Hospital and Postgraduate Institute of Ophthalmology, Tamil Nadu, Dr. Y. C. Venugopal Reddy, Chief, Aravind Eye Hospital and Postgraduate Institute of Ophthalmology, Tamil Nadu
Project Overview
To determine the utility of various polymerase chain reaction (PCR) methods in intra-ocular fluids for detecting the infectious genomes in the diagnosis of infectious endophthalmitis in terms of sensitivity and specificity. This prospective and consecutive analysis included a total of 326 intra-ocular fluids and four other ocular specimens that were submitted for the microbiological evaluation, which were obtained from 330 clinically diagnosed endophthalmitis patients presented between October 2012 and September 2015 at the tertiary eye care referral centre in South India. Part of the collected intra-ocular fluids were subjected to cultures and smears, and the remaining parts were utilized for five PCR uniplex, nested, semi-nested, multiplex and nested multiplex after extracting DNA, using universal eubacterial and Propionibacterium acnes speciesspecific primer sets targeting 16S rRNA gene in all bacteria and pan-fungal primers, targeting 28S rRNA gene in all fungi.
Professor, Aravind Eye Hospital and Postgraduate Institute of Ophthalmology, Tamil Nadu
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