Molecular detection of<i>Paracoccidioides brasiliensis</i>in soil

Theodoro, Raquel Cordeiro; Candeias, J. A. N.; Araújo, João Pessoa; Bosco, Sandra de Moraes Gimenes; Macoris, Severino Assis Da Graça; Padula, Luis Otavio; Franco, Marcello; Bagagli, Eduardo

doi:10.1080/13693780500129418

Cited by 58 publications

(45 citation statements)

References 27 publications

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“…The limit of detection for the OTsn-PCR assay was comparable with that of the protocol developed by Koishi et al 5 ( Figure 1B) and was more sensitive than the protocols in the Imai et al 8 and Theodoro et al 9 studies (Figures 1C-1D). Furthermore, when using DNA extracted from the clinical samples, the OTsn-PCR assay and the protocol developed by Koishi et al 5 were able to detect P. brasiliensis DNA in all of the samples (100%) ( Table 1), while the protocols of Imai et al 8 and Theodoro et al 9 only detected P. brasiliensis DNA in four (28.5%) and eight (57.2%) of the clinical samples, respectively (Table 1). A possible cause for this difference may be the lower numbers of cycles used in these reactions.…”

supporting

confidence: 54%

“…This OTsn-PCR approach was compared with the nested and semi-nested PCR methods described by Imai et al 8 , Theodoro et al 9 and Koishi et al 5 Similar to our OTsn-PCR assay, these reactions use sequences of ribosomal DNA as targets, especially from the ITS, and are specifi c for the detection of P. brasiliensis. For the sake of comparison, we used fi ve different serially diluted P. brasiliensis DNA concentrations (2.5ng, 250pg, 25pg, 2.5pg and 0.25pg).…”

mentioning

confidence: 99%

“…All of the protocols were repeated at least three times, as originally described. Furthermore, to compare cycle number effects on the limit of detection among the methods, the methods developed by Imai et al 8 and Theodoro et al 9 were also performed using 35 cycles.…”

mentioning

confidence: 99%

“…A possible cause for this difference may be the lower numbers of cycles used in these reactions. For this reason, all of samples were retested following the Imai et al 8 and Theodoro et al 9 PCR methods using 35 cycles. The numbers of positive samples increased from four to six (42.8%) and from eight to 10 (71.4%) using the PCR methods of Imai et al 8 and Theodoro et al 9 , respectively (Table 1).…”

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confidence: 99%

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An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection

Pitz

Koishi

Tavares

et al. 2013

Rev. Soc. Bras. Med. Trop.

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Revista da Sociedade Brasileira de Medicina Tropical 46(6):783-785, Nov-Dec, 2013http://dx.doi. org/10.1590/0037-8682-1625-2013 An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection ABSTRACT Introduction: Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis. Methods: We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods. Results: The OTsn-PCR assay was positive for all clinical samples, and the detection limit was better or equivalent to the other nested or semi-nested PCR methods for P. brasiliensis detection. Conclusions:The OTsn-PCR assay described in this paper has a detection limit similar to other reactions for the molecular detection of P. brasiliensis, but this approach is faster and less prone to contamination than other conventional nested or semi-nested PCR assays.

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confidence: 54%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection

Pitz

Koishi

Tavares

et al. 2013

Rev. Soc. Bras. Med. Trop.

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“…The presence of P. brasiliensis in soil samples collected from armadillo burrows could be confirmed molecularly, by applying a species-specific Nested PCR, based on ITS-5.8S-ITS2 regions [69]. It is interesting to note that the pathogen could be detected mainly in soil samples collected in the interior of the armadillo burrows [70].…”

Section: Origin and Maintenance Of P Brasiliensis' Virulencementioning

confidence: 93%

Paracoccidioides brasiliensis: phylogenetic and ecological aspects

et al. 2008

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The knowledge on the biological aspects of Paracoccidioides brasiliensis has evolved greatly since the first description of the disease in 1908. From the pioneers, who were able to clearly demonstrate the fungal nature of the agent, to the recent genomic era, important advances have been achieved. P. brasiliensis is a true fungus, belonging to the Ascomycetous Division, although its sexual phase has not been demonstrated morphologically. A better understanding of the fundamental aspects of the agent, especially its phylogeny and evolutionary history, will provide us with valuable insights allowing a better comprehension of the disease and our capacity to deal with the problem. Concerning the fungus's ecology, although some progress had been observed, the ecological niche of the pathogen has not been determined yet. The aim of the present review is to focus on the biological aspects of P. brasiliensis from an evolutionary point of view, addressing the fungus's phylogenetic aspects, in those special points that might be relevant for the pathogen/ host interactions, the biological forces that have been acting on its origin and maintenance of virulence, as well as in determining the fungus's ecology and epidemiology.

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Fundamentals of paracoccidioidomycosis treatment

Fróes

Caligiorne

2011

Drug Development Research

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Paracoccidioidomycosis (PCM) is the most important mycosis in Latin America. The etiological agent is the fungus, Paracoccidioides brasiliensis. Approximately 1 million people are infected with the fungus, 10% of whom will show symptoms of the disease. However, clinical and laboratorial studies addressing the diagnosis for this disease are both scarce and diverse in terms of definition. Nevertheless, two main advances were included in the portfolio of available diagnostic methods for paracoccidioidomycosis during the last decades: the detection of antibodies against P. brasiliensis antigens by serological methods, and the detection of the fungi DNA in peripheral blood and fragments of lesions using the polymerase chain reaction (PCR) assay and species-specific primers. Normally, the treatment of PCM is quite prolonged because of the immunosuppressed profile of the patients and the ability of P. brasiliensis to survive in the tissues even after long-term treatment. Case reports indicate that azoles and sulfa compounds can produce positive results in less severe cases of PCM. However, for acute cases of the disease in immunocompromised patients or in cases of neuroparacoccidioidomycosis, there is no standard treatment. Thus, there is a need to establish a standard treatment protocol within the context of the patient clinical profile. Moreover, drug availability to public institutions is an important issue, since new drugs, e.g., second-generation azoles, the echinocandines and immunomodulatory drugs, are not yet available in public hospitals and clinics. Drug Dev Res 72:528-537, 2011.

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Molecular detection ofParacoccidioides brasiliensisin soil

Cited by 58 publications

References 27 publications

An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection

An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection

Paracoccidioides brasiliensis: phylogenetic and ecological aspects

Fundamentals of paracoccidioidomycosis treatment

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