Premium
Assessment of a pan‐dermatophyte nested‐ PCR compared with conventional methods for direct detection and identification of dermatophytosis agents in animals
Author(s) -
Piri Fahimeh,
Zarei Mahmoudabadi Ali,
Ronagh Ali,
Ahmadi Bahram,
Makimura Koichi,
RezaeiMatehkolaei Ali
Publication year - 2018
Publication title -
mycoses
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.13
H-Index - 69
eISSN - 1439-0507
pISSN - 0933-7407
DOI - 10.1111/myc.12821
Subject(s) - dermatophyte , nested polymerase chain reaction , polymerase chain reaction , biology , primer (cosmetics) , trichophyton , microbiology and biotechnology , genetics , chemistry , gene , antifungal , organic chemistry
Summary 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 DNA s 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.