A new PCR primer for the identification of Paracoccidioides brasiliensis based on rRNA sequences coding the internal transcribed spacers (ITS) and 5·8S regions

T. Imai Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, Chuo-ku, Chiba (260-8673), Japan

A. Sano Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, Chuo-ku, Chiba (260-8673), Japan

Y. Mikami Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, Chuo-ku, Chiba (260-8673), JapanCorrespondence[email protected]

K. Watanabe Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, Chuo-ku, Chiba (260-8673), Japan

F. H. Aoki Faculty of Medical Science, the State University of Campinas, São Paulo, Brazil

M.L. M. Branchini Faculty of Medical Science, the State University of Campinas, São Paulo, Brazil

R. Negroni Facultad de Medicina-UBA, Centro de Micologia, Departamento de Microbiologia, Paraguay 2155 Piso 11, Buenos Aires, Argentina

K. Nishimura Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, Chuo-ku, Chiba (260-8673), Japan

M. Miyaji Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, Chuo-ku, Chiba (260-8673), Japan

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Abstract

Internal transcribed spacer (ITS) genes including the 5·8S ribosomal (r)RNA of Paracoccidioides brasiliensis were amplified and the DNA sequences were determined. Based on a comparison of the sequence information, a new polymerase chain reaction (PCR) primer pair was designed for specific amplification of DNA for P. brasiliensis. This primer pair amplified a 418-bp DNA sequence and was 100% successful in identifying 29 strains of P. brasiliensis (including the reference strains) isolated from the regions of Brazil, Costa Rica, Japan, Argentina or from different sources. The results of specificity tests of these primers to compare the fungus with those of Aspergillus fumigatus, Blastomyces dermatitidis, Candida albicans, Cryptococcus neoformans, Histoplasma capsulatum and Penicillium marneffei are also reported.

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A new PCR primer for the identification of Paracoccidioides brasiliensis based on rRNA sequences coding the internal transcribed spacers (ITS) and 5·8S regions

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A new PCR primer for the identification of Paracoccidioides brasiliensis based on rRNA sequences coding the internal transcribed spacers (ITS) and 5·8S regions

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