1.Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis.
Alfredo GARCIA ; Remigio MARTINEZ ; Jose Manuel BENITEZ-MEDINA ; David RISCO ; Waldo Luis GARCIA ; Joaquin REY ; Juan Manuel ALONSO ; Javier Hermoso DE MENDOZA
Journal of Veterinary Science 2013;14(4):491-494
Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was used to evaluate clinical samples collected from naturally infected animals with D. congolensis. The results showed that this assay is a fast and reliable method for diagnosing dermatophilosis.
Actinomycetales/*isolation & purification
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Actinomycetales Infections/diagnosis/microbiology/*veterinary
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Animals
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Cattle
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Cattle Diseases/*diagnosis/microbiology
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Fluorescent Dyes/*diagnostic use
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Horse Diseases/*diagnosis/microbiology
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Horses
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Limit of Detection
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Real-Time Polymerase Chain Reaction/*methods/veterinary
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Reproducibility of Results
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Sheep
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Sheep Diseases/*diagnosis/microbiology