Role of Sanger Sequencing in the Early Diagnosis of Infective Endophthalmitis: Experience From a Pilot Study.

Background This study aimed to evaluate the role of Sanger sequencing in the early diagnosis of infectious endophthalmitis and compare its diagnostic yield with conventional microbiological methods. Methodology A prospective pilot study was conducted at a tertiary eye care center over a period of one year, including 11 patients with clinically suspected endophthalmitis. Vitreous samples were subjected to Gram staining, KOH wet mount, and aerobic/fungal culture. Parallel molecular testing was performed using 16S rRNA PCR and Sanger sequencing. Diagnostic yield, turnaround time, and clinical relevance were compared. Results Gram stain and KOH wet mount were positive in 1/11 (9.1%) cases each, while cultures were sterile in 9/11 (81.8%) and showed contaminants in 2/11 (18.2%). PCR amplification succeeded in 6/11 (54.5%) samples, with Sanger sequencing identifying Achromobacter (one case) and Bacillus spp. (three cases). Overall, molecular diagnostics established a likely bacterial etiology in 6/11 (54.5%) patients. Sequencing provided results within 24-48 hours, versus 5-7 days for culture. Conclusions Sanger sequencing significantly improved diagnostic yield and reduced turnaround time compared with conventional techniques. Although not a replacement for culture, it represents a valuable adjunct in managing culture-negative endophthalmitis. Larger multicentric studies are warranted to validate its routine clinical role.