Minglan Shi1, Xiqing Li1, Jiao Feng1, Shulin Jia2, Xing Xiao1, Chunmei Chen1, Cindy Fransisca1, Liyan Xi1, Junmin Zhang1. 1. Department of Dermatology and Venerology, Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. 2. Department of Dermatology, Guangdong Province Hospital of Chinese Medicine, Guangzhou, China.
Abstract
BACKGROUND: Chromoblastomycosis (CBM) is a chronic fungal disease. In China, the principle etiologic agent was a group of dematiaceous fungi, including Fonsecaea monophora, Fonsecaea nubica, and Cladophialophora carrionii. Although the Fonsecaea species have similar morphology, their pathogenicity is quite different. This study aims to establish a new solution for early identification of Fonsecaea species because of their distinctive potential infection risk. METHODS: Five reference strains and 35 clinical isolates from patients with CBM, preserved in our laboratory, were used in this study. The universal primer ITS1 and ITS2 were chosen to amplify the highly conserved regions of rDNA. High-resolution melting (HRM) analysis was performed using the LIGHTCYCLER® 96 System. All the amplicons were verified by direct sequencing and the sequence were aligned with those in GenBank by BLAST analysis. RESULTS: We successfully differentiated the five strains according to their different Tm values and curve shapes. The 35 clinical isolates from patients were identified as 24 strains for F. monophora and 11 strains for F. nubica, which is consistent with the DNA sequencing results. CONCLUSION: It is the first time to use HRM analysis for identification of Fonsecaea species. Since the CBM etiologic agent in South China is mainly F. monophora and F. nubica, this strategy is sufficient to be applied in the clinical examination with high accuracy, speed, and throughput.
BACKGROUND:Chromoblastomycosis (CBM) is a chronic fungal disease. In China, the principle etiologic agent was a group of dematiaceous fungi, including Fonsecaea monophora, Fonsecaea nubica, and Cladophialophora carrionii. Although the Fonsecaea species have similar morphology, their pathogenicity is quite different. This study aims to establish a new solution for early identification of Fonsecaea species because of their distinctive potential infection risk. METHODS: Five reference strains and 35 clinical isolates from patients with CBM, preserved in our laboratory, were used in this study. The universal primer ITS1 and ITS2 were chosen to amplify the highly conserved regions of rDNA. High-resolution melting (HRM) analysis was performed using the LIGHTCYCLER® 96 System. All the amplicons were verified by direct sequencing and the sequence were aligned with those in GenBank by BLAST analysis. RESULTS: We successfully differentiated the five strains according to their different Tm values and curve shapes. The 35 clinical isolates from patients were identified as 24 strains for F. monophora and 11 strains for F. nubica, which is consistent with the DNA sequencing results. CONCLUSION: It is the first time to use HRM analysis for identification of Fonsecaea species. Since the CBM etiologic agent in South China is mainly F. monophora and F. nubica, this strategy is sufficient to be applied in the clinical examination with high accuracy, speed, and throughput.
Authors: Sara Gago; Óscar Zaragoza; Isabel Cuesta; Juan L Rodríguez-Tudela; Manuel Cuenca-Estrella; María J Buitrago Journal: J Clin Microbiol Date: 2011-08-10 Impact factor: 5.948
Authors: Amanda Bombassaro; Sybren de Hoog; Vinicius A Weiss; Emanuel M Souza; Aniele C R Leão; Flávia F Costa; Valter Baura; Michele Z Tadra-Sfeir; Eduardo Balsanelli; Leandro F Moreno; Roberto T Raittz; Maria Berenice R Steffens; Fabio O Pedrosa; Jiufeng Sun; Liyan Xi; Anamélia L Bocca; Maria S Felipe; Marcus Teixeira; Germana D Santos; Flávio Q Telles Filho; Conceição M P S Azevedo; Renata R Gomes; Vânia A Vicente Journal: Genome Announc Date: 2016-07-28