PCR-AGE, automated and manual methods to identify Candida strains from veterinary sources: a comparative approach.
The increasing incidence of candidiasis has drawn the attention of scientists and clinicians attempting to improve methods of studying Candida yeasts. PCR amplification followed by agarose gel electrophoresis (PCR-AGE) and the manual method (morphological characteristics, biochemical profiles and culturing on CHROMagar-Candida) and VITEK 2 automated method were used to test a total of 30 fungal strains from dog sources. The strains were obtained from cases of dermatitis, otitis externa and from the ears, oral mucosa, vaginal mucosa, prepuce and perianal region of clinically normal dogs. After identification as Candida yeasts by the manual method, the strains were analyzed using both VITEK and PCR-AGE methods. Isolates of C. parapsilosis ATCC 22019, C. krusei ATCC 6258 and C. albicans ATCC 10231 were included as controls. The universal primers ITS1, ITS3 and ITS4 were used in two independent PCR reactions. Of 30 yeast isolates, 3 isolates (Saccharomyces cerevisiae, C. rugosa and C. parapsilosis) that were incompletely identified by the manual method were identified with the PCR-AGE and VITEK methods. The results revealed a 96.7% and 86.7% concurrent identification between the PCR-AGE and VITEK methods versus the manual method, respectively. PCR-AGE showed a greater level of concordance with the manual method, besides being faster and more sensitive than the other methods examined, and is therefore indicated for routine diagnostic testing of Candida spp. strains from veterinary sources.
Brito
Erika H S
EH
Faculty of Veterinary, Postgraduate Program in Veterinary Science, State University of Ceará, Fortaleza-CE, Ceará, Brazil.
Brilhante
Raimunda S N
RS
Cordeiro
Rossana A
RA
Sidrim
José J C
JJ
Fontenelle
Raquel O S
RO
Melo
Luciana M
LM
Albuquerque
Erica S
ES
Rocha
Marcos F G
MF
D000818:Animals; D001331:Automation / Q000379:methods; D002175:Candida / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D002177:Candidiasis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D004285:Dogs; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005260:Female; D016533:Mycological Typing Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer