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Validation of PCR for the detection of pseudomonas aeruginosa from cornea specimens
Author(s) -
KOWALSKI R,
HILLENBRAND M,
THOMPSON P
Publication year - 2010
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2010.4451.x
Subject(s) - pseudomonas aeruginosa , microbiology and biotechnology , cornea , 16s ribosomal rna , primer (cosmetics) , keratitis , biology , pseudomonas , bacteria , chemistry , genetics , organic chemistry , neuroscience
Purpose The definitive detection of Pseudomonas aeruginosa from the cornea is essential for effective therapy especially in cases complicated by anti‐infective pre‐treatment. 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]. Conclusion 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.