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16S Ribosomal RNA PCR Versus Conventional Diagnostic Culture in the Investigation of Suspected Bacterial Keratitis

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Date 2020 Dec 21
PMID 33344046
Citations 11
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Abstract

Purpose: The purpose of this study was to compare conventional diagnostic culture (CDC) to 16S ribosomal RNA polymerase chain reaction (PCR) analysis for diagnosing bacterial keratitis.

Methods: Samples collected from 100 consecutive patients presenting to the Royal Liverpool University Hospital with bacterial keratitis were processed using CDC and 16S PCR analysis.

Results: The overall detection rate using both methods was 36%. Of these, 72.2% (26/36) were detected by PCR and 63.9% (23/36) isolated by CDC ( = 0.62). Using a combination of both PCR and CDC increased the detection rate for pathogenic bacteria by 13% compared to using CDC alone ( = 0.04). In CDC negative samples, 16S PCR identified more pathogens than CDC in 16S PCR negative samples. Neither order of sample collection nor prior antimicrobial use affected the detection rate.

Conclusions: 16S rRNA gene PCR performed in addition to CDC on corneal samples from patients with clinically suspected bacterial keratitis led to additional pathogen detection.

Translational Relevance: 16S rRNA gene PCR should be developed to become an additional part of clinical service for patients with bacterial keratitis rather than used in isolation.

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References
1.
Bottger E . Rapid determination of bacterial ribosomal RNA sequences by direct sequencing of enzymatically amplified DNA. FEMS Microbiol Lett. 1989; 53(1-2):171-6. DOI: 10.1016/0378-1097(89)90386-8. View

2.
Balne P, Reddy A, Kodiganti M, Gorli S, Garg P . Evaluation of three PCR assays for the detection of fungi in patients with mycotic keratitis. Br J Ophthalmol. 2012; 96(6):911-2. DOI: 10.1136/bjophthalmol-2011-300207. View

3.
Kim E, Chidambaram J, Srinivasan M, Lalitha P, Wee D, Lietman T . Prospective comparison of microbial culture and polymerase chain reaction in the diagnosis of corneal ulcer. Am J Ophthalmol. 2008; 146(5):714-23, 723.e1. DOI: 10.1016/j.ajo.2008.06.009. View

4.
Itahashi M, Higaki S, Fukuda M, Shimomura Y . Detection and quantification of pathogenic bacteria and fungi using real-time polymerase chain reaction by cycling probe in patients with corneal ulcer. Arch Ophthalmol. 2010; 128(5):535-40. DOI: 10.1001/archophthalmol.2010.66. View

5.
Bourcier T, Thomas F, Borderie V, Chaumeil C, Laroche L . Bacterial keratitis: predisposing factors, clinical and microbiological review of 300 cases. Br J Ophthalmol. 2003; 87(7):834-8. PMC: 1771775. DOI: 10.1136/bjo.87.7.834. View