Molecular identification of Nocardia species causing endophthalmitis using multilocus sequence analysis (MLSA): a 10-year perspective
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Updated: Dec 23, 2025

Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
Published on: July 11, 2016
Hariharan Gnanam1, Siva Ganesa Karthikeyan Rajapandian2, Rameshkumar Gunasekaran2
1Department of Biotechnology, Alagappa University, Karaikudi, India.
Abstract:
Introduction. Nocardia spp. can cause several ocular infections, such as keratitis, endophthalmitis and scleral abscesses. Molecular identification of Nocardia spp. by 16S rDNA sequencing is the gold standard method at present for species identification, but closely related species can only be identified by multilocus sequence analysis (MLSA) of housekeeping genes.Aim. The major objective was to profile Nocardia species, antibiotic susceptibility patterns and clinical outcomes in endophthalmitis patients.Methodology. Between January 2009 and December 2018, endophthalmitis patients who were diagnosed with Nocardia infection based on microscopic and culture characteristics were selected. Antibacterial susceptibility tests were performed and Nocardia speciation was performed using MLSA and phylogenetic tree analysis of the 16 s rRNA gene and the gyrB, hsp65 and secA1 genes.Results. A total of 43 culture-proven patients were identified during the study period. All isolates were 100 % sensitive to amikacin and 98 % resistant to ceftazidime. Fluoroquinolone sensitivity was observed in the range of 58 to 72 %. Year-wise analysis of antibiotic resistance patterns revealed there was a significant increase in resistance to fluoroquinolones. Twenty-two isolates were stored and six different species were identified. Nocardia farcinica (n=10) was found to be the most predominant, followed by Nocardia cyriacigeorgica (n=4), Nocardia otitidiscaviarum (n=3), Nocardia amikacinitolerans (n=2), Nocardia puris (n=2) and Nocardia higoensis (n=1).Conclusions. N. farcinica is the major pathogen, and this is the first report to identify N. otitidiscaviarum, N. amikacinitolerans and N. higoensis as causing endophthalmitis. Overall, visual outcomes were mostly poor even after aggressive management.
Insights
Nocardia endophthalmitis is primarily caused by Nocardia farcinica, with increasing fluoroquinolone resistance observed. This study identifies new Nocardia species causing endophthalmitis, highlighting poor visual outcomes despite treatment.
Area of Science:
- Ophthalmology
- Medical Microbiology
- Infectious Diseases
Background:
- Nocardia species are known to cause severe ocular infections, including endophthalmitis.
- Accurate species identification is crucial for effective treatment, with multilocus sequence analysis (MLSA) being superior to 16S rDNA sequencing for closely related species.
Purpose of the Study:
- To characterize Nocardia species causing endophthalmitis.
- To determine antibiotic susceptibility patterns of Nocardia isolates.
- To evaluate clinical outcomes in patients with Nocardia endophthalmitis.
Main Methods:
- Retrospective analysis of 43 endophthalmitis patients with Nocardia infection (January 2009-December 2018).
- Antibacterial susceptibility testing was performed.
- Nocardia speciation utilized MLSA, including 16S rRNA, gyrB, hsp65, and secA1 genes.
Main Results:
- Nocardia farcinica was the most common species (n=10), followed by N. cyriacigeorgica (n=4), N. otitidiscaviarum (n=3), N. amikacinitolerans (n=2), N. puris (n=2), and N. higoensis (n=1).
- All isolates were susceptible to amikacin, while 98% were resistant to ceftazidime.
- Fluoroquinolone sensitivity ranged from 58-72%, with a significant increase in resistance over time.
Conclusions:
- Nocardia farcinica is the predominant cause of endophthalmitis in this cohort.
- This study reports Nocardia otitidiscaviarum, Nocardia amikacinitolerans, and Nocardia higoensis as causative agents of endophthalmitis for the first time.
- Visual outcomes for Nocardia endophthalmitis remain poor despite aggressive management.
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