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

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.

Comprehensive Molecular Dissection of Dermatophilus congolensis Genome and First Observation of tet(Z) Tetracycline Resistance

“…Rarely, the gene clusters such as bacteriocins, siderophores, and terpenes were found, however, their homology to similar proteins in other bacterial species was very low. This random occurrence of bacteriocins, siderophores, and terpenes may suggest their horizontal pattern of acquisition as reported for other bacterial species [34].…”

RT‐qPCR for the diagnosis of dermatophilosis in horses

“…Conventional PCR tests and a SYBR green PCR assay have been described . Conventional PCR is time‐consuming and involves an increased number of steps in order to definitively identify a specific organism.…”

“…Molecular diagnostics allow for the rapid and accurate detection of infectious microorganisms. Conventional and SYBR green based assays have been described for the detection of D. congolensis . The purpose of this study was to develop a Taqman real time quantitative PCR (RT‐qPCR) test for the detection of D. congolensis and validate this with clinical cases.…”

Identification and Antimicrobial Resistance of Dermatophilus congolensis from Cattle in Saint Kitts and Nevis

“…After purification, preliminary identification of D. congolensis was performed based on the phenotypic appearance and classical biochemical reactions of indole and catalase. Identification was confirmed by PCR, as described [ 11 ] with slight modifications. Genomic bacterial DNA was purified using the Qiagen Allprep bacterial DNA/RNA/protein kit (Qiagen, Hilden, Germany.…”