J Vet Sci.  2013 Dec;14(4):491-494. 10.4142/jvs.2013.14.4.491.

Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis

• Garcia A
• Martinez R
• Benitez-Medina JM
• Risco D
• Garcia WL
• Rey J
• Alonso JM
• de Mendoza JH

• KMID: 1712318
• DOI: http://doi.org/10.4142/jvs.2013.14.4.491

Abstract

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.

Keyword

clinical samples; Dermatophilus congolensis; dermatophilosis; diagnosis; real-time PCR

MeSH Terms

Actinomycetales/*isolation & purification
Actinomycetales Infections/diagnosis/microbiology/*veterinary
Animals
Cattle
Cattle Diseases/*diagnosis/microbiology
Fluorescent Dyes/*diagnostic use
Horse Diseases/*diagnosis/microbiology
Horses
Limit of Detection
Real-Time Polymerase Chain Reaction/*methods/veterinary
Reproducibility of Results
Sheep
Sheep Diseases/*diagnosis/microbiology
Fluorescent Dyes

Figure

• 

  Fig. 1 (A) The linear standard curve obtained for the dilution series of D. congolensis DNA ranging from 10 ng to 1.10-4 ng. (B) Real time (RT)-PCR results for the clinical samples.

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