RESUMO
Background: Histoplasma capsulatum and Pneumocystis spp. may cause a host infection through the respiratory airway, mainlyaffecting the pulmonary tissue. These fungal pathogens affect a wide range of mammalian species, including humans and bats.The co-infection of bats with both organisms above has never been studied in Brazil. The aim of the present research was todetect the presence of the H. capsulatum and Pneumocystis spp. in lung samples of bat species from two states of Brazil. For thispurpose, a highly sensitive nested PCR was used with specific molecular markers for each pathogen.Materials, Methods & Results: Two hundred and forty-nine bats were captured between 2007 and 2009 in caves, forests, andurban areas of Mato Grosso (MT) and Rio Grande do Sul (RS), located respectively in the Mid-Western and Southern regions. Thebats were captured following the guidelines of the rabies control manual for herbivores, standardized by the Ministry of the Agriculture. Detection of Pneumocystis spp. DNA was based upon nested PCR, which amplified a portion of the mitochondrial smallsubunit (mtSSU) of the rRNA gene, whereas the H. capsulatum DNA was amplified employing the Hcp 100 locus. Amplificationproducts were sequenced to confirm fungal presence in bat lungs. The amplifications results for H. capsulatum and Pneumocystisspp. were positive in 63 [25.3%, IC95% (20.1%-31.25%)] and 95 [(38,2%, IC95% (32.1%-44.52%)] samples, respectively. Thegreatest occurrence of Histoplasma capsulatum was observed in Desmodus rotundus (20.6%), Tadarida brasiliensis (20.6%),Histiotus velatus (19.0%) and Molossus molossus (11.1%), with the detection in the other species being lower than 7.9%, amongthe 24 studied bat species. For Pneumocystis spp., the detection was higher in Tadarida brasiliensis (23.1%), Desmodus rotundus(18.%), Histiotus velatus (14.7%), and Molossus molossus (11,6%), being lower than 5.3% in the other species. A co-infection...(AU)
Assuntos
Animais , Quirópteros/microbiologia , Pulmão/microbiologia , Histoplasma/isolamento & purificação , Pneumocystis/isolamento & purificação , Coinfecção/veterinária , Doenças Respiratórias/microbiologia , BrasilRESUMO
Background: Histoplasma capsulatum and Pneumocystis spp. may cause a host infection through the respiratory airway, mainlyaffecting the pulmonary tissue. These fungal pathogens affect a wide range of mammalian species, including humans and bats.The co-infection of bats with both organisms above has never been studied in Brazil. The aim of the present research was todetect the presence of the H. capsulatum and Pneumocystis spp. in lung samples of bat species from two states of Brazil. For thispurpose, a highly sensitive nested PCR was used with specific molecular markers for each pathogen.Materials, Methods & Results: Two hundred and forty-nine bats were captured between 2007 and 2009 in caves, forests, andurban areas of Mato Grosso (MT) and Rio Grande do Sul (RS), located respectively in the Mid-Western and Southern regions. Thebats were captured following the guidelines of the rabies control manual for herbivores, standardized by the Ministry of the Agriculture. Detection of Pneumocystis spp. DNA was based upon nested PCR, which amplified a portion of the mitochondrial smallsubunit (mtSSU) of the rRNA gene, whereas the H. capsulatum DNA was amplified employing the Hcp 100 locus. Amplificationproducts were sequenced to confirm fungal presence in bat lungs. The amplifications results for H. capsulatum and Pneumocystisspp. were positive in 63 [25.3%, IC95% (20.1%-31.25%)] and 95 [(38,2%, IC95% (32.1%-44.52%)] samples, respectively. Thegreatest occurrence of Histoplasma capsulatum was observed in Desmodus rotundus (20.6%), Tadarida brasiliensis (20.6%),Histiotus velatus (19.0%) and Molossus molossus (11.1%), with the detection in the other species being lower than 7.9%, amongthe 24 studied bat species. For Pneumocystis spp., the detection was higher in Tadarida brasiliensis (23.1%), Desmodus rotundus(18.%), Histiotus velatus (14.7%), and Molossus molossus (11,6%), being lower than 5.3% in the other species. A co-infection...
Assuntos
Animais , Coinfecção/veterinária , Histoplasma/isolamento & purificação , Pneumocystis/isolamento & purificação , Pulmão/microbiologia , Quirópteros/microbiologia , Brasil , Doenças Respiratórias/microbiologiaRESUMO
A high prevalence of Pneumocystis jirovecii colonization was observed in patients positive for the human immunodeficiency virus (HIV) admitted to a tertiary hospital in southern Brazil between August 2012 and December 2012. Amplification of the mitochondrial large subunit ribosomal RNA gene in oropharyngeal samples through nested polymerase chain reaction identified P. jirovecii colonization in 26 of 58 (44.8%) HIV-positive patients admitted for causes other than Pneumocystis pneumonia. Colonization was more frequent among patients with an absolute CD4 count ≤200 cells/µl. These findings suggest that the HIV-infected population is a major reservoir and source of P. jirovecii infection and that identification of such individuals may contribute to future strategies for improving management of HIV-infected patients.
Assuntos
Infecções Oportunistas Relacionadas com a AIDS/epidemiologia , Portador Sadio/epidemiologia , Pneumocystis carinii , Pneumonia por Pneumocystis/epidemiologia , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Adulto , Brasil/epidemiologia , Portador Sadio/microbiologia , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Orofaringe/microbiologia , Pneumocystis carinii/genética , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , PrevalênciaRESUMO
BACKGROUND: Neurosyphilis diagnosis is frequently dependent upon the results of serological tests and cerebrospinal fluid abnormalities, but the reliability of findings in patients with HIV-1 infection has been questioned, especially in asymptomatic patients with latent syphilis. In this study, we present the data on the presence of T. pallidum DNA in CSF from asymptomatic HIV-infected patients with the diagnosis of syphilis. METHODS: CSF and serum samples were collected from 12 HIV-infected patients attending a tertiary care clinic located in southern Brazil, during the period 2012 to 2013. RESULTS: In CSF samples from five of 12 patients (40%), we detected T. pallidum DNA. Unexpectedly, in these patients, the CSF cell count, protein and glucose levels were normal. In addition, none of these 5 CSF samples presented a positive VDRL reaction. Serum VDRL titers were similar between patients with positive and negative CSF T. pallidum DNA. Most patients with detectable T. pallidum DNA presented low serum VDRL titers. A higher serum VDRL titer of 1:64 was observed in only one patient. CONCLUSIONS: Our results have shown that asymptomatic HIV-infected patients with evidence of latent syphilis and normal CSF might present detectable T. pallidum DNA in the CSF. The detection of T. pallidum DNA by our seminested PCR provides additional information beyond conventional CSF analysis for the diagnosis of neurosyphilis. The detection of T. pallidum DNA in CSF despite normal CSF findings in HIV-infected patients could also provide a different therapeutic approach including the use of intravenous aqueous crystalline penicillin.
Assuntos
Líquido Cefalorraquidiano/microbiologia , Coinfecção , DNA Bacteriano/genética , Infecções por HIV/complicações , Neurossífilis/diagnóstico , Reação em Cadeia da Polimerase/métodos , Sífilis Latente/diagnóstico , Treponema pallidum/genética , Adolescente , Adulto , Doenças Assintomáticas , Brasil , DNA Bacteriano/líquido cefalorraquidiano , Feminino , Infecções por HIV/diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade , Neurossífilis/líquido cefalorraquidiano , Neurossífilis/microbiologia , Valor Preditivo dos Testes , Sífilis Latente/líquido cefalorraquidiano , Sífilis Latente/microbiologia , Centros de Atenção Terciária , Fatores de TempoRESUMO
A high rate of Pneumocystis jirovecii colonization was observed in Brazilian cystic fibrosis (CF) patients (13 out of 34; 38.2%) who underwent bronchoscopy between March 2006 and August 2009 at the Hospital de Clinicas de Porto Alegre, Brazil. Bronchoalveolar lavage samples were collected from these patients and studied by nested PCR amplification of the mitochondrial gene coding for the large subunit ribosomal RNA (mtLSUrDNA). The observed rate of colonization was higher than that reported in European populations. Genotypic characterization of the mtLSUrDNA locus revealed a predominance of the polymorphisms 85C/248C (genotype 1) and 85T/248C (genotype 3), with all samples possessing the wild-type genotype of dihydropteroate synthase. These findings suggest that cystic fibrosis patients could be an important reservoir and source of P. jirovecii infection. Further studies are required to elucidate the role of this common fungal colonization in the evolution of CF patients.
Assuntos
Fibrose Cística/microbiologia , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/epidemiologia , Adolescente , Adulto , Brasil/epidemiologia , Criança , Pré-Escolar , Fibrose Cística/complicações , Fibrose Cística/epidemiologia , DNA Fúngico/genética , DNA Mitocondrial/genética , DNA Ribossômico/genética , Feminino , Genes de RNAr , Humanos , Lactente , Masculino , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/complicações , Pneumonia por Pneumocystis/microbiologia , Reação em Cadeia da Polimerase , Prevalência , Análise de Sequência de DNA , Adulto JovemRESUMO
Background: Pneumocystis constitutes a highly diversified biological group, with numerous species, which are strongly host-specific and well adapted to live inside the lungs of a diverse range of mammals. The detection of DNA from Pneumocystis in clinical specimens by PCR assays is leading to important advances in pneumonia caused by Pneumocystis and its epidemiology. The aim of this study was to analyze two different diagnostic methods, real-time PCR (qPCR) using primers based in the Major Surface Glycoprotein (MSG) of Pneumocystis sp. and conventional nested PCR using primers designed to the small subunit of mitochondrial rRNA (mtSSU rRNA) for detection of Pneumocystis DNA in lung tissue from bats. Materials, Methods & Results: Bats (195 samples) were captured (2007-2009) in caves, forests, and urban areas, were obtained from the Program of Rabies Control of two states in Brazil: Mato Grosso and Rio Grande do Sul, located respectively in the Mid-Western and Southern regions of the country approximately 2000 km apart. Lung tissue (250 mg) was finely minced, homogenized with crushing and DNA extraction was carried out with commercial kit. DNA samples the lung tissue of bats were analyzed by nested PCR, using oligonucleotide primers designed for the gene encoding the mitochondrial small subunit rRNA (mtSSU rRNA) and Taqman probe and primers for qPCR were selected based on the Major Surface Glycoprotein (MSG) of Pneumocystis sp. Chi-square (P < 0.001 was considered signifi cant) and the McNemar's test was used to analyze nested PCR and qPCR as methods of detection of Pneumocystis sp. and the Kappa was calculated by Win Episcope 2.0. To assess the sensitivity and specificity of the qPCR assay, a nested PCR assay was considered as the reference method. The positivity was 36.4% in the nested PCR and 24.1% using the qPCR. Concordance was obtained in 68.2% of the samples (133/195). It was demonstrated that there was no statistically significant difference between the techniques used and, both tests proved to be specific for the detection of Pneumocystis species. Specificity was 71% for the nested PCR and 84.6% for the qPCR. Pneumocystis was detected (71/195) by the nested PCR assay in 14 species:. Tadarida brasiliensis, Histiotus velatus, Desmodus rotundus, Molossus molossus, Glossophaga soricina, Nyctinomops laticaudatus, Promops nasutus, Artibeus sp., Eptesocus furinalus, Lasurus blossevillii, Molossus currentium, Molossus rufus, Myotis levis and Nyctinomops macrotis. Discussion: This study detected the DNA from Pneumocystis through the nested PCR and qPCR assays, and the frequency found is comparable to that obtained in a previous study, which used the nested PCR in Central American, South American and European countries. Pneumocystis sp. was observed in a high number of different bat species (14) in two Brazilian States (RS and MT). The qPCR showed a higher specificity in comparison to the nested PCR. The literature has similar findings to the results obtained by this research, employing the same tests and genes. The nested PCR and qPCR assays are indicated in the diagnosis of Pneumocystis sp. in bats and it is important to highlight that a better diagnostic precision is achieved with the association of both tests. Additionally, this study was the first to detect Pneumocystis sp. in the lungs of bats using qPCR.
Assuntos
Animais , Pneumocystis/isolamento & purificação , Pneumonia por Pneumocystis/veterinária , Quirópteros , Reação em Cadeia da Polimerase/veterinária , Genes de RNAr , Reação em Cadeia da Polimerase em Tempo Real/veterinária , PulmãoRESUMO
Background: Pneumocystis constitutes a highly diversifi ed biological group, with numerous species, which are strongly host-specifi c and well adapted to live inside the lungs of a diverse range of mammals. The detection of DNA from Pneumocystis in clinical specimens by PCR assays is leading to important advances in pneumonia caused by Pneumocystis and its epidemiology. The aim of this study was to analyze two different diagnostic methods, real-time PCR (qPCR) using primers based in the Major Surface Glycoprotein (MSG) of Pneumocystis sp. and conventional nested PCR using primers designed to the small subunit of mitochondrial rRNA (mtSSU rRNA) for detection of Pneumocystis DNA in lung tissue from bats.Materials, Methods & Results: Bats (195 samples) were captured (2007-2009) in caves, forests, and urban areas, were obtained from the Program of Rabies Control of two states in Brazil: Mato Grosso and Rio Grande do Sul, located respectively in the Mid-Western and Southern regions of the country approximately 2000 km apart. Lung tissue (250 mg) was fi nely minced, homogenized with crushing and DNA extraction was carried out with commercial kit. DNA samples the lung tissue of bats were analyzed by nested PCR, using oligonucleotide primers designed for the gene encoding the mitochondrial small subunit r RNA (mtSSU rRNA) and Taqman probe and primers for qPCR were selected based on
Background: Pneumocystis constitutes a highly diversifi ed biological group, with numerous species, which are strongly host-specifi c and well adapted to live inside the lungs of a diverse range of mammals. The detection of DNA from Pneumocystis in clinical specimens by PCR assays is leading to important advances in pneumonia caused by Pneumocystis and its epidemiology. The aim of this study was to analyze two different diagnostic methods, real-time PCR (qPCR) using primers based in the Major Surface Glycoprotein (MSG) of Pneumocystis sp. and conventional nested PCR using primers designed to the small subunit of mitochondrial rRNA (mtSSU rRNA) for detection of Pneumocystis DNA in lung tissue from bats.Materials, Methods & Results: Bats (195 samples) were captured (2007-2009) in caves, forests, and urban areas, were obtained from the Program of Rabies Control of two states in Brazil: Mato Grosso and Rio Grande do Sul, located respectively in the Mid-Western and Southern regions of the country approximately 2000 km apart. Lung tissue (250 mg) was fi nely minced, homogenized with crushing and DNA extraction was carried out with commercial kit. DNA samples the lung tissue of bats were analyzed by nested PCR, using oligonucleotide primers designed for the gene encoding the mitochondrial small subunit r RNA (mtSSU rRNA) and Taqman probe and primers for qPCR were selected based on
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Background: Pneumocystis constitutes a highly diversifi ed biological group, with numerous species, which are strongly host-specifi c and well adapted to live inside the lungs of a diverse range of mammals. The detection of DNA from Pneumocystis in clinical specimens by PCR assays is leading to important advances in pneumonia caused by Pneumocystis and its epidemiology. The aim of this study was to analyze two different diagnostic methods, real-time PCR (qPCR) using primers based in the Major Surface Glycoprotein (MSG) of Pneumocystis sp. and conventional nested PCR using primers designed to the small subunit of mitochondrial rRNA (mtSSU rRNA) for detection of Pneumocystis DNA in lung tissue from bats.Materials, Methods & Results: Bats (195 samples) were captured (2007-2009) in caves, forests, and urban areas, were obtained from the Program of Rabies Control of two states in Brazil: Mato Grosso and Rio Grande do Sul, located respectively in the Mid-Western and Southern regions of the country approximately 2000 km apart. Lung tissue (250 mg) was fi nely minced, homogenized with crushing and DNA extraction was carried out with commercial kit. DNA samples the lung tissue of bats were analyzed by nested PCR, using oligonucleotide primers designed for the gene encoding the mitochondrial small subunit r RNA (mtSSU rRNA) and Taqman probe and primers for qPCR were selected based on
Background: Pneumocystis constitutes a highly diversifi ed biological group, with numerous species, which are strongly host-specifi c and well adapted to live inside the lungs of a diverse range of mammals. The detection of DNA from Pneumocystis in clinical specimens by PCR assays is leading to important advances in pneumonia caused by Pneumocystis and its epidemiology. The aim of this study was to analyze two different diagnostic methods, real-time PCR (qPCR) using primers based in the Major Surface Glycoprotein (MSG) of Pneumocystis sp. and conventional nested PCR using primers designed to the small subunit of mitochondrial rRNA (mtSSU rRNA) for detection of Pneumocystis DNA in lung tissue from bats.Materials, Methods & Results: Bats (195 samples) were captured (2007-2009) in caves, forests, and urban areas, were obtained from the Program of Rabies Control of two states in Brazil: Mato Grosso and Rio Grande do Sul, located respectively in the Mid-Western and Southern regions of the country approximately 2000 km apart. Lung tissue (250 mg) was fi nely minced, homogenized with crushing and DNA extraction was carried out with commercial kit. DNA samples the lung tissue of bats were analyzed by nested PCR, using oligonucleotide primers designed for the gene encoding the mitochondrial small subunit r RNA (mtSSU rRNA) and Taqman probe and primers for qPCR were selected based on
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BACKGROUND: Infliximab, a chimeric antitumour necrosis factor (TNF) monoclonal antibody, has become an established effective therapy for inflammatory rheumatic disease. However, TNF is a critical factor in host defence, and the suppression of its biological activity may be associated with the increased risk of opportunistic infections. The frequent use of infliximab in clinical practice has identified Pneumocystis jirovecii pneumonia (PcP) as a serious complication. Individuals colonized with Pneumocystis may be at high risk of development of PcP when they have undergone immunosuppression. Hence, we addressed the question of the frequency of Pneumocystis colonization among patients treated with infliximab. DESIGN: We examined 125 oropharyngeal washes collected from 78 individuals with rheumatoid arthritis, 30 with ankylosing spondylitis and 17 with psoriatic arthritis, half of them underwent infliximab therapy, using a real-time polymerase chain reaction assay that employs specific primers from a portion of the mitochondrial large-subunit rRNA gene of P. jirovecii. RESULTS: Pneumocystis jirovecii colonization was detected in 32 (25·6%) patients. In a multivariate regression model, only duration of infliximab treatment for more than 3 years and use of corticosteroid were significantly and independently associated with risk of Pneumocystis colonization. However, the effect of corticosteroid on P. jirovecii colonization rate was not linearly dose dependent as showed other logistic regression analysis. CONCLUSIONS: There is a high rate of P. jirovecii colonization among patients with rheumatologic diseases treated with infliximab. The identification of patients colonized by P. jirovecii before starting the treatment with infliximab could be a strategy for PcP prevention.
Assuntos
Anti-Inflamatórios/uso terapêutico , Anticorpos Monoclonais/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Infecções Oportunistas/induzido quimicamente , Pneumonia por Pneumocystis/induzido quimicamente , Espondiloartropatias/tratamento farmacológico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Hospedeiro Imunocomprometido , Infliximab , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Infecções Oportunistas/microbiologia , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , Adulto JovemRESUMO
Pneumocystis jirovecii, the fungal agent that causes Pneumocystis pneumonia (PCP), is known to exclusively infect humans. Molecular studies have enabled detection of this fungus in individuals who have been colonized by P. jirovecii. Such colonization, found in several populations, seems to act as a human reservoir for the fungus. Various studies have reported mutations associated with sulfa resistance in P. jirovecii strains isolated from colonized patients, who can transmit the mutant genotype to PCP-susceptible individuals. The growing interest in P. jirovecii colonization may prompt the design of new prevention and management strategies for PCP.
Assuntos
Portador Sadio/epidemiologia , Reservatórios de Doenças/microbiologia , Pneumocystis carinii , Pneumonia por Pneumocystis/epidemiologia , Infecções Oportunistas Relacionadas com a AIDS/epidemiologia , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Animais , Antifúngicos , Portador Sadio/microbiologia , Farmacorresistência Fúngica Múltipla/genética , Previsões , Genótipo , Humanos , Mamíferos/microbiologia , Mutação de Sentido Incorreto , Pneumocystis carinii/efeitos dos fármacos , Pneumocystis carinii/genética , Pneumocystis carinii/isolamento & purificação , Pneumocystis carinii/patogenicidade , Pneumonia por Pneumocystis/transmissão , Especificidade da EspécieRESUMO
Several studies from developed countries have documented the association between trimethoprim-sulfamethoxazole prophylaxis failure and mutations in the Pneumocystis jirovecii gene coding for dihydropteroate synthase (DHPS). DNA was extracted from Giemsa-stained smears of 70 patients with P. jirovecii pneumonia seen in Porto Alegre, Brazil, from 1997 to 2004. Successful PCR amplification of the DHPS locus was obtained in 57 of 70 cases (81.4%), including five cases (8.7%) that had used sulfa prophylaxis. No DHPS gene mutations were seen. These results suggest that DHPS mutations are currently as rare in Brazil as in other developing countries.