title: Molecular identification of Histoplasma capsulatum using rolling circle amplification.
abstract: Histoplasmosis is a systemic fungal disease that occurs worldwide, causing symptomatic infection mostly in immunocompromised hosts. Etiological agent is the dimorphic fungus, Histoplasma capsulatum, which occurs in soil contaminated with bird or bat droppings. Major limitation in recognition of H. capsulatum infections is the low awareness, since other diseases may have similar symptomatology. The molecular methods have gained importance because of unambiguous diagnostic ability and efficiency. The aim of this study was to develop and evaluate a padlock probe in view of rolling circle amplification (RCA) detection method which targets ITS (Internal Transcribed Spacer) rDNA of H. capsulatum enabling rapid and specific detection of the fungus in clinical samples. Two padlock probes were designed and one of these (HcPL2) allowed specific amplification of H. capsulatum DNA while no cross-reactivity was observed with fungi used as negative controls. This method proved to be effective for H. capsulatum specific identification and demonstrated to be faster than the traditional method of microbiological identification.
authors.affiliation: Bioprocess Engineering and Biotechnology Department, Post-Graduation Program of Bioprocess Engineering and Biotechnology, Federal University of Paraná, Curitiba, Brazil.
authors.forename: Jason L
authors.initials: JL
authors.lastname: Furuie
authors.affiliation: Guangdong Provincial Institute of Public Health, Guangdong Provincial Center for Disease Control and Prevention, Guangzhou, China.
authors.forename: Jiufeng
authors.initials: J
authors.lastname: Sun
authors.affiliation: Basic Pathology Department, Post-Graduation Program of Microbiology, Parasitology and Pathology, Federal University of Paraná, Curitiba, Brazil.
authors.forename: Mariana M F
authors.initials: MM
authors.lastname: do Nascimento
authors.affiliation: Basic Pathology Department, Post-Graduation Program of Microbiology, Parasitology and Pathology, Federal University of Paraná, Curitiba, Brazil.
authors.forename: Renata R
authors.initials: RR
authors.lastname: Gomes
authors.affiliation: Basic Pathology Department, Post-Graduation Program of Microbiology, Parasitology and Pathology, Federal University of Paraná, Curitiba, Brazil.
authors.forename: Caroline E
authors.initials: CE
authors.lastname: Waculicz-Andrade
authors.affiliation: Ecossistema Environmental Consulting, Curitiba, Brazil.
authors.forename: Gisele C
authors.initials: GC
authors.lastname: Sessegolo
authors.affiliation: Cellular Biology Division, Microbiology, Immunology and Parasitology Department, Federal University of São Paulo, São Paulo, Brazil.
authors.forename: Anderson M
authors.identifier: http://orcid.org/0000-0003-0891-4563
authors.initials: AM
authors.lastname: Rodrigues
authors.affiliation: Center for Zoonosis Control, São Paulo, Brazil.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Galvão-Dias
authors.affiliation: Cellular Biology Division, Microbiology, Immunology and Parasitology Department, Federal University of São Paulo, São Paulo, Brazil.
authors.forename: Zoilo P
authors.initials: ZP
authors.lastname: de Camargo
authors.affiliation: Basic Pathology Department, Post-Graduation Program of Microbiology, Parasitology and Pathology, Federal University of Paraná, Curitiba, Brazil.|Clinical Hospital of the Federal University of Paraná, Curitiba, Brazil.
authors.forename: Flávio
authors.initials: F
authors.lastname: Queiroz-Telles
authors.affiliation: Parasitology and Mycology Department, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Mohammad J
authors.initials: MJ
authors.lastname: Najafzadeh
authors.affiliation: Basic Pathology Department, Post-Graduation Program of Microbiology, Parasitology and Pathology, Federal University of Paraná, Curitiba, Brazil.|CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands.
authors.forename: Sybren G
authors.initials: SG
authors.lastname: de Hoog
authors.affiliation: Bioprocess Engineering and Biotechnology Department, Post-Graduation Program of Bioprocess Engineering and Biotechnology, Federal University of Paraná, Curitiba, Brazil.|Basic Pathology Department, Post-Graduation Program of Microbiology, Parasitology and Pathology, Federal University of Paraná, Curitiba, Brazil.
authors.forename: Vania A
authors.initials: VA
authors.lastname: Vicente
mesh_terms: D000818:Animals; D015342:DNA Probes; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D006658:Histoplasma / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D006660:Histoplasmosis / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D010802:Phylogeny; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer
keywords: Histoplasma capsulatum; Histoplasmosis; RCA; diagnosis; padlock probe