Assessment of a pan-dermatophyte nested-PCR compared with conventional methods for direct detection and identification of dermatophytosis agents in animals
| Autor: | Koichi Makimura, Fahimeh Piri, Ali Rezaei-Matehkolaei, Ali Zarei Mahmoudabadi, Bahram Ahmadi, Ali Ronagh |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
030106 microbiology Dermatology Commercial kit Biology medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity Microbiology Animal Diseases 030207 dermatology & venereal diseases 03 medical and health sciences 0302 clinical medicine Dogs Tinea Translation elongation medicine Animals Horses DNA Fungal Microscopy Sheep Arthrodermataceae Goats Direct microscopy General Medicine Infectious Diseases Dermatophyte Cats Cattle Nested polymerase chain reaction Animal skin |
| Zdroj: | Mycoses. 61(11) |
| ISSN: | 1439-0507 |
| Popis: | Conventional direct microscopy with potassium hydroxide (KOH) and culture were found to lack the ability to establish a fast and specific diagnosis of dermatophytosis. A pan-dermatophyte nested-PCR assay was developed using a novel primer pair targeting the translation elongation factor 1-α (Tef-1α) sequences for direct detection and identification of most veterinary relevant dermatophytes in animal samples suspected to dermatophytosis. A total of 140 animal skin and hair samples were subjected to direct microscopy, culture, and ITS-RFLP/ITS-sequencing of culture isolates for the detection and identification of dermatophytosis agents. Nested-PCR sequencing was performed on all the extracted DNAs using a commercial kit after dissolving the specimens by mechanical beating. Nested-PCR was positive in 90% of samples, followed by direct microscopy (85.7%) and culture (75%). The degree of agreement between nested-PCR and direct microscopy (94.4%) was higher than with culture (83.3%). In 105 culture-positive cases, the measures of agreement for the identification of dermatophytosis agents were as follows: 100% between nested-PCR sequencing and ITS-RFLP/ITS-sequencing and 63.8% between nested-PCR sequencing and culture. The developed nested-PCR was faster as well as more sensitive and specific than conventional methods for detection and identification of dermatophytes in clinical samples, which was particularly suitable for epidemiological studies. |
| Databáze: | OpenAIRE |
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