Please use this identifier to cite or link to this item:
https://www.arca.fiocruz.br/handle/icict/58088
Type
ArticleCopyright
Restricted access
Embargo date
2028-12-31
Collections
- INI - Artigos de Periódicos [3460]
Metadata
Show full item record
HYPERBRANCHED ROLLING CIRCLE AMPLIFICATION AS A RAPID AND SENSITIVE METHOD FOR SPECIES IDENTIFICATION WITHIN THE CRYPTOCOCCUS SPECIES COMPLEX
Cryptococcus gattii
Hyperbranched rolling circle amplification
Internal transcribed spacer
Species identification
Affilliation
Molecular Mycology Research Laboratory. Centre for Infectious Diseases and Microbiology. Westmead Millennium Institute. Westmead Hospital. Westmead, Australia / Inter-Department of Medical. Microbiology Graduate School. Chulalongkorn University. Bangkok, Thailand.
Retroviral Genetics Laboratory. Centre for Virus Research. Westmead Millennium Institute. Westmead Hospital. The University of Sydney. Sydney, Australia.
Molecular Mycology Research Laboratory. Centre for Infectious Diseases and Microbiology. Westmead Millennium Institute. Westmead Hospital. Westmead, Australia.
Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brazil.
Centre for Infectious Diseases and Microbiology. Public Health, Institute of Clinical Pathology and Medical Research. Westmead Hospital. Westmead, Australia.
Molecular Mycology Research Laboratory. Centre for Infectious Diseases and Microbiology. Westmead Millennium Institute. Westmead Hospital. Westmead, Australia / Faculty of Medicine. The University of Sydney. Western Clinical School at Westmead Hospital. Sydney, Australia.
Retroviral Genetics Laboratory. Centre for Virus Research. Westmead Millennium Institute. Westmead Hospital. The University of Sydney. Sydney, Australia.
Molecular Mycology Research Laboratory. Centre for Infectious Diseases and Microbiology. Westmead Millennium Institute. Westmead Hospital. Westmead, Australia.
Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brazil.
Centre for Infectious Diseases and Microbiology. Public Health, Institute of Clinical Pathology and Medical Research. Westmead Hospital. Westmead, Australia.
Molecular Mycology Research Laboratory. Centre for Infectious Diseases and Microbiology. Westmead Millennium Institute. Westmead Hospital. Westmead, Australia / Faculty of Medicine. The University of Sydney. Western Clinical School at Westmead Hospital. Sydney, Australia.
Abstract
The Cryptococcus species complex contains two closely related basidiomycetous yeasts: Cryptococcus neoformans and C. gattii, which cause cryptococcosis in humans and other animals. The species and varieties are characterized, by different clinical, epidemiological, biochemical and molecular features. The currently used identification methods are either time-consuming or not anymore commercially available. However, a rapid, sensitive and robust assay for the detection of these pathogens is vital for early diagnosis and appropriate treatment decisions. To overcome those limitations, four padlock probes targeting species-specific single nucleotide polymorphisms at the internal transcribed spacers (ITSs) of the RNA gene locus were developed and applied during isothermal hyperbranched rolling circle amplification (HRCA). The probes were tested against 99 samples, including 94 clinical cryptococcal cultures, three closely related Cryptococcus species, and two clinical specimens. The use of the padlock probes and the combination of probe signal amplification by HRCA provided a quick and sensitive assay for the accurate identification of C. neoformans var. grubii, C. neoformans var. neoformans and C. gattii. HRCA was also useful to detect hybrids, when they were heterozygous at the ITS locus. The HRCA results were in agreement with previous genotyping data based on PCR fingerprinting, amplified fragment length polymorphism and ITS sequencing.
Keywords
Cryptococcus neoformansCryptococcus gattii
Hyperbranched rolling circle amplification
Internal transcribed spacer
Species identification
Share