PCR-Restriction Fragment Length Polymorphism Analysis of the Phospholipase B (
            <i>PLB1</i>
            ) Gene for Subtyping of
            <i>Cryptococcus neoformans</i>
            Isolates

Latouche, GN; Huynh, Matthew; Sorrell, Tania C.; Meyer, Wieland

doi:10.1128/aem.69.4.2080-2086.2003

Cited by 56 publications

(57 citation statements)

References 38 publications

(39 reference statements)

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“…In the present work, VNI was prevalent in 45/47 isolates (95.7%) while VNII was 2/47 (4.3%), none new molecular subtype was found. Our results enhance previous data that suggest the differences based on more conserved genes as PLB1 or URA-5 by PCR-RFLP can be resultant of ongoing divergent evolution within the C. neoformans complex, into the current eight subtypes and not only coincidental 32 .…”

Section: Discussionsupporting

confidence: 78%

“…The PCR-RFLP method based on PLB1 gene is of great importance because highlights differences based on partial sequence of a single virulence gene, while PCR-fingerprinting, on the other hand, amplifies mostly noncoding DNA sequences across the whole genome 14,32 . In our knowledge, there are not reports evolving Brazilian samples using this method.…”

Section: Discussionmentioning

confidence: 99%

“…The PLB1 fragment was amplified by PCR, in a final volume of 50 µL, digested with AvaI in accordance with LATOUCHE et al 32 and was separated by electrophoresis on 3% agarose gel at 100V for 5.5 hours.…”

Section: Pcr-rflp Analysis Of the Plb1 Genementioning

confidence: 99%

“…Several molecular typing methods have been used in epidemiological analyses of clinical and/or environmental isolates of C. neoformans, including karyotyping, PCR fingerprinting, random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) and amplified fragment length polymorphism (AFLP) 5,7,8,10,12,16,22,26,37,45 . PCR-fingerprinting using mini (M13) or microsatellite [(GACA) 4 ] specific sequences as single primers and RFLP (PLB1 and URA5 genes) methods have grouped global isolates of C. neoformans into eight major molecular types 32,37,38 . Likely, BOEKHOUT et al using AFLP analysis grouped into six major AFLP groups 8 .…”

Section: Introductionmentioning

confidence: 99%

“…The phospholipase B (PLB1) gene of C. neoformans has been described as important virulence factor in a mouse and rabbit and as interesting target for molecular typing of C. neoformans via PCR-RFLP analysis 15,32 . However until the present moment, there are not available data about the molecular epidemiology of C. neoformans using PLB1-RFLP method in Brazil.…”

Section: Introductionmentioning

confidence: 99%

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Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil

Matsumoto

Fusco‐Almeida

Baeza

et al. 2007

Rev. Inst. Med. trop. S. Paulo

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SUMMARYThe basidiomycetous yeast Cryptococcus neoformans is an important fungal pathogen mainly in immunocompromised patients. In this study, 47 clinical isolates of C. neoformans from regions of São Paulo State were studied serologically by using the Crypto Check Iatron RM 304-K kit, their genetic diversity was estimated by PCR-fingerprinting with a microsatellite-specific sequence (GACA) 4 , RAPD with primer 6 (Amersham Pharmacia Biotech), PCR-restriction fragment length polymorphism (RFLP) analysis of the phospholipase B gene (PLB1) digested with AvaI and mating type analysis by PCR. All 47 strains isolated from HIV positive patients included in this study were serotype A and MATα. The majority of the isolates (45/47) were VNI and only two were VNII by PCR-fingerprinting and PCR-RFLP analysis. High degree of homogeneity was observed when (GACA) 4 was used, being highly correlated (> 0.9). In contrast, the RAPD analysis was more heterogeneous with higher number of molecular profiles. By PCR-RFLP, no new molecular type was found, enhancing the suggestion that the differences based on conserved gene as PLB1, can be resultant of ongoing divergent evolution within the C. neoformans complex, into the current eight subtypes. Our results furnish new information on the molecular epidemiology of C. neoformans in the southeast region of Brazil.

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Section: Discussionsupporting

confidence: 78%

Section: Discussionmentioning

confidence: 99%

Section: Pcr-rflp Analysis Of the Plb1 Genementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil

Matsumoto

Fusco‐Almeida

Baeza

et al. 2007

Rev. Inst. Med. trop. S. Paulo

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Hyperbranched rolling circle amplification as a rapid and sensitive method for species identification within the Cryptococcus species complex

et al. 2008

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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 speciesspecific 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.

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Current awareness on yeast

2003

Yeast

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In order to keep subscribers up‐to‐date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly‐published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (4 weeks journals ‐ search completed 19th. Feb. 2003)

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PCR-Restriction Fragment Length Polymorphism Analysis of the Phospholipase B ( PLB1 ) Gene for Subtyping of Cryptococcus neoformans Isolates

Abstract: Cryptococcus neoformans is a pathogenic yeast that is currently divided into three varieties

Cited by 56 publications

References 38 publications

Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil

Genotyping, serotyping and determination of mating-type of Cryptococcus neoformans clinical isolates from São Paulo State, Brazil

Hyperbranched rolling circle amplification as a rapid and sensitive method for species identification within the Cryptococcus species complex

Current awareness on yeast

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