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2010 OMIG, Abstract 18

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Validation of PCR for the detection of Pseudomonas aeruginosa from cornea specimens
R. P. Kowalski, M.A. Hillenbrand, E.G. Romanowski, P.P. Thompson,
The Charles T. Campbell Ophthalmic Microbiology Laboratory, The University of Pittsburgh, Pittsburgh, PA

Purpose: The definitive detection of Pseudomonas aeruginosa from the cornea is essential for effective therapy.
Methods: The present study investigated the validation of PCR for detecting Pseudomonas aeruginosa from corneal specimens using a real-time method with the SmartCycler (Cepheid, Sunnyvale, CA, USA). Primer and probes directed toward the ecfx (63 bp) and the 16s rRNA genes were validated against 20 true-positive specimens (corneal specimens positive for Pseudomonas aeruginosa) and 20 true-negative samples (spiked samples of HSV, VZV, Candida, Aspergillus, and Fusarium).
Results: The sensitivity, specificity, positive predictive value, negative predictive value, and efficiency of PCR to the ecfx and 16s rDNA genes were, respectively: [80%, 95%, 95%, 80%, 87%], and [77%, 100%, 100%, 77%, 87%]. The amplification efficacy and limit of detection were respectively: [0.97, 33.6 µg/ml], and [1.03, 8.12 µg/ml].
Conclusions: PCR was validated for detecting Pseudomonas aeruginosa from corneal specimens. The impact of detection of Pseudomonas aeruginosa from cases of non-specified keratitis will need to be determined.

Support: Pennsylvania Lion’s Club, USA
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