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1.
J Med Microbiol ; 68(11): 1649-1654, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31609198

RESUMO

Myeloid C-type lectin receptors (CLRs) are innate immune recognition molecules that bind to microorganisms via their carbohydrate recognition domains. In this study, we utilized a library of CLRs that recognize fungal mannans. We used this library to screen against Pneumocystis carinii (Pc) homogenates or purified Pc major surface glycoprotein (Msg) present on Pneumocystis. The results demonstrated that all of the mammalian CLR hFc-fusions tested displayed significant interaction/binding with Pc organisms, and furthermore to isolated Msg. Highest Pc organism and Msg binding activities were with CLR members Mincle, Dectin-2, DC-SIGN and MCL. An immunofluorescence assay with the respective CLR hFc-fusions against whole Pc life forms corroborated these findings. Although some of these CLRs have been implicated previously as important for Pneumocystis pathogenesis (Dectin-1/Dectin-2/Mincle), this is the first analysis of head-to-head comparison of known fungal mannan binding CLR-hFc fusions with Pc. Lastly, heat treatment resulted in reducted CLR binding.


Assuntos
Proteínas Fúngicas/metabolismo , Lectinas Tipo C/metabolismo , Mananas/metabolismo , Glicoproteínas de Membrana/metabolismo , Infecções por Pneumocystis/metabolismo , Pneumocystis carinii/metabolismo , Humanos , Lectinas Tipo C/genética , Infecções por Pneumocystis/genética , Infecções por Pneumocystis/microbiologia , Pneumocystis carinii/genética , Ligação Proteica
2.
Gac Med Mex ; 155(4): 377-385, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31486786

RESUMO

Introduction: Pneumocystis jirovecii is an atypical fungus particularly detected in HIV-positive or transplant patients. Objective: To detect and genotype Pneumocystis jirovecii in patient samples from two hospitals in Mexico City. Method: Eighty-nine respiratory tract samples, corresponding to 53 patients (30 HIV-positive and 23 HIV-negative) with respiratory symptoms and to 11 healthy individuals included as negative control, were processed. DNA was extracted from the ITS region and amplified by nested polymerase chain reaction from the internal transcribed spacer, with one fragment being obtained at each round (693 and 550 bp). Genotypes and their phylogenetic relationship were determined by sequencing the 550 bp fragment. Results: Forty-eight samples from 30 HIV-positive patients were received from a single hospital, out of which 11 (36.6 %) were positive for Pneumocystis jirovecii. No sample was positive in HIV-negative patients or healthy subjects. The most frequently detected haplotypes were Eg and Em. Conclusions: The frequency of Pneumocystis jirovecii infection was high in the studied Mexican population. The most common genotype was different from those reported in other countries. It is necessary to address this health problem through early detection of this infection.


Assuntos
Infecções por HIV/complicações , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/epidemiologia , Adulto , Idoso , Pré-Escolar , Estudos Transversais , Feminino , Genótipo , Humanos , Masculino , México , Pessoa de Meia-Idade , Filogenia , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/diagnóstico , Pneumonia por Pneumocystis/microbiologia , Reação em Cadeia da Polimerase , Estudos Prospectivos , Adulto Jovem
3.
Braz J Infect Dis ; 23(5): 352-357, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31545952

RESUMO

Exposure to Pneumocystis jirovecii (P. jirovecii) can lead to a wide variety of presenting features ranging from colonization in immunocompetent patients with lung disease, to invasive infections in immunocompromised hosts. Colonization by this fungus in patients with chronic obstructive pulmonary disease (COPD) could be associated with higher rates of exacerbations and impaired lung function in these patients. Our objective was to determine whether colonization by P. jirovecii in patients with COPD is associated with increased exacerbations and deterioration of lung function. This was a prospective cohort study on patients with COPD. All participants meeting selection criteria underwent clinical and microbiological assessments and were then classified as colonized vs. non-colonized patients. Chi-squared tests were performed and multivariate logistic models were fitted in order to obtain risk ratios (RR) with 95% confidence intervals (CI). We documented a frequency of colonization by P. jirovecii of 32.3%. Most patients were categorized as having GOLD B and D COPD. The history of significant exacerbations in the last year, health status impairment (COPD Assesment Tool ≥10), airflow limitation (percent of post-bronchodilator FEV1), and BODEx score (≥5) were similar between groups. After a 52-week follow-up period, the rate of adjusted significant exacerbations did not differ between groups. However, a decrease in FEVI was found in both groups.


Assuntos
Pulmão/fisiopatologia , Infecções por Pneumocystis/microbiologia , Pneumocystis carinii/genética , Doença Pulmonar Obstrutiva Crônica/microbiologia , Idoso , Progressão da Doença , Feminino , Humanos , Masculino , Infecções por Pneumocystis/fisiopatologia , Pneumocystis carinii/isolamento & purificação , Estudos Prospectivos , Doença Pulmonar Obstrutiva Crônica/fisiopatologia
4.
Diagn Cytopathol ; 47(11): 1194-1196, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31322837

RESUMO

Liver transplant recipients are prone to several infections, including lung infections, which can lead to substantial morbidity and mortality. Bronchoalveolar lavage (BAL) cytology is a rapid and sensitive diagnostic tool to identify the etiologic agents. We report a rare case of a 24-year-old male, post Live donor liver transplantation for autoimmune chronic liver disease, who presented with cough, fever, weight loss, and cavitatory lesion in lung. BAL cytology revealed Leishmania donovani (LD) and Pneumocystis jirovecii/carinii (PCP). Cytomegalovirus deoxyribonucleic acid polymerase chain reaction (CMV DNA PCR) test showed markedly raised levels. Patient was put on treatment for these multiple infections and showed significant improvement. Thus, rapid diagnosis of infections through BAL cytology is crucial in transplant recipients to institute timely therapy and avoid undesirable empirical treatments. Moreover, this case highlights a rare finding of LD bodies along with PCP in BAL cytology.


Assuntos
Líquido da Lavagem Broncoalveolar , Lavagem Broncoalveolar , Infecções por Citomegalovirus , Leishmania donovani/genética , Leishmaniose Visceral , Transplante de Fígado , Pneumocystis carinii/genética , Pneumonia por Pneumocystis , Reação em Cadeia da Polimerase , Adulto , Líquido da Lavagem Broncoalveolar/microbiologia , Líquido da Lavagem Broncoalveolar/parasitologia , Líquido da Lavagem Broncoalveolar/virologia , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/genética , Infecções por Citomegalovirus/patologia , Humanos , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/genética , Leishmaniose Visceral/patologia , Doadores Vivos , Masculino , Pneumonia por Pneumocystis/diagnóstico , Pneumonia por Pneumocystis/genética , Pneumonia por Pneumocystis/patologia
5.
BMC Infect Dis ; 19(1): 658, 2019 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-31337356

RESUMO

BACKGROUND: Pneumocystis jirovecii pneumonia (PCP) is one of the most common HIV-related opportunistic infections. The diagnosis of PCP is based on analyses from respiratory tract specimens which may require the invasive procedure of a diagnostic bronchoscopy. The objective of this study was to evaluate the diagnostic potential of Pneumocystis jirovecii PCR in serum combined with the 1,3-ß-D-glucan (betaglucan) test for the diagnosis of PCP in HIV-infected patients. METHODS: This was a retrospective case-control study including serum samples from 26 HIV-infected patients with PCP collected within 5 days prior to the start of PCP treatment, 21 HIV-infected control subjects matched by blood CD4+ cell counts, and 18 blood donors. The serum samples were analyzed for Pneumocystis jirovecii PCR and betaglucan. The reference standard for PCP was based on previously described microbiological and clinical criteria. RESULTS: All patients with PCP had detectabe Pneumocystis jirovecii DNA in serum yielding a sensitivity for the Pneumocystis jirovecii PCR assay in serum of 100%. All blood donors had negative Pneumocystis PCR in serum. The specificity when testing HIV-infected patients was 71%, but with a PCR Cycle threshold (Ct) value of 34 as cut-off the specificity was 90%. At a putative pretest probaility of 20%, the negative and positive predictive value for the Pneumocystis PCR assay in serum was 0.99 and 0.71, respectively. Betaglucan with cut-off level 200 pg/ml combined with a positive Pneumocystis jirovecii PCR result had sensitivity and specificity of 92 and 90%, respectively. The concentration of Pneumocystis jirovecii DNA in serum samples, expressed by the PCR Ct values, correlated inversely to the betaglucan levels in serum. CONCLUSION: In this case-control study including 70% of all HIV-infected patients with PCP treated at Sahlgrenska University Hospital during a time period of 13 years, Pneumocystis PCR analysis on serum samples had a very high sensitivity and negative predictive value for the diagnosis of PCP in HIV-infected patients. A serum-based diagnostic procedure either based on Pneumocystis jirovecii PCR alone or in combination with betaglucan analysis may thus be feasible and would facilitate the care of HIV-infected patients with suspected PCP.


Assuntos
Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/microbiologia , beta-Glucanas/sangue , Adolescente , Adulto , Idoso , Doadores de Sangue , Estudos de Casos e Controles , DNA Fúngico/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/patogenicidade , Pneumonia por Pneumocystis/sangue , Reação em Cadeia da Polimerase , Estudos Retrospectivos , Sensibilidade e Especificidade
6.
Int J Infect Dis ; 86: 65-72, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31207386

RESUMO

BACKGROUND: Histoplasma capsulatum and Pneumocystis jirovecii are respiratory fungal pathogens that principally cause pulmonary disease. Coinfection with both pathogens is scarcely reported. This study detected this coinfection using specific molecular methods for each fungus in the bronchoalveolar lavage (BAL) of patients from a tertiary care hospital. MATERIALS AND METHODS: BAL samples from 289 hospitalized patients were screened by PCR with specific markers for H. capsulatum (Hcp100) and P. jirovecii (mtLSUrRNA and mtSSUrRNA). The presence of these pathogens was confirmed by the generated sequences for each marker. The clinical and laboratory data for the patients were analyzed using statistical software. RESULTS: The PCR findings separated three groups of patients, where the first was represented by 60 (20.8%) histoplasmosis patients, the second by 45 (15.6%) patients with pneumocystosis, and the last group by 12 (4.2%) patients with coinfection. High similarity among the generated sequences of each species was demonstrated by BLASTn and neighbor-joining algorithms. The estimated prevalence of H. capsulatum and P. jirovecii coinfection was higher in HIV patients.


Assuntos
Coinfecção/epidemiologia , Histoplasmose/epidemiologia , Pneumocystis carinii , Pneumonia por Pneumocystis/epidemiologia , Adulto , Idoso , Lavagem Broncoalveolar , Feminino , Infecções por HIV/complicações , Histoplasma/genética , Histoplasma/isolamento & purificação , Histoplasmose/microbiologia , Humanos , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Pneumocystis carinii/genética , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , Reação em Cadeia da Polimerase , Centros de Atenção Terciária
7.
J Mycol Med ; 29(2): 107-111, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31047784

RESUMO

Pneumocystis pneumonia (PCP) is a potentially life-threatening fungal infection usually seen in immunocompromised patients. Pneumocystis jirovecii can be easily detected from oral rinse samples in HIV patients with suspected PCP. In this study, a quantitative real-time PCR assay was used to establish the frequency of detection of P. jirovecii in oral rinses from HIV patients without respiratory symptoms or suspicion of PCP. Two saline oral rinses were collected from 100 ambulant HIV patients and from 60 COPD patients (comparator group). Four HIV patients were positive for P. jirovecii. In three patients, the first sample was positive and in one the second one was positive. One of these patients was on PCP prophylaxis and had a CD4+ count of 76 cells/mm3. The mean CD4+ count for all patients was 527 cells/mm3. All qRT-PCR test results for the COPD patients were negative. No patient developed PCP at six months follow-up. The qRT-PCR assay can be used to detect P. jirovecii DNA in oral rinse samples from HIV patients without evident clinical symptoms, however the oral carriage of this fungus was rare in our cohort of patients. In conclusion, although rare, a positive oral rinse P. jirovecii result may reflect colonisation, in particular in patients with HIV. This needs to be kept in mind when using oral rinses and qRT-PCR in the diagnosis of P. jirovecii infection.


Assuntos
Infecções Assintomáticas , Infecções por HIV/complicações , Boca/microbiologia , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Contagem de Linfócito CD4 , Estudos de Coortes , DNA Fúngico/genética , Feminino , Infecções por HIV/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/genética , Reação em Cadeia da Polimerase em Tempo Real , Solução Salina , Reino Unido , Adulto Jovem
9.
J Infect Chemother ; 25(6): 477-479, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30797688

RESUMO

Detecting Pneumocystis jirovecii by bronchoalveolar lavage or lung biopsy is the gold standard for diagnosis of P. jirovecii pneumonia (PJP); however, these techniques are not always applicable in children because of their high invasiveness. We report two pediatric cases of PJP diagnosed by polymerase chain reaction (PCR) of gastric lavage that were successfully treated. To date, there are no reported cases of using PCR of gastric lavage to diagnose PJP. On the day of PJP onset, both the infants required respiratory support and infiltrative shadows were observed in both lung fields on chest radiography. Furthermore, their (1 â†’ 3)-ß-D glucan levels were elevated. P. jirovecii was detected by PCR of gastric lavage and trimethoprim-sulfamethoxazole was administered for 3 weeks, following which their condition improved. They were long-term steroid users, but without any prophylaxis. PCR of gastric lavage in cases of suspected PJP may help in confirming the diagnosis in children who have mild to moderate airway symptoms, or have difficulty with invasive examination like bronchoscopy.


Assuntos
Lavagem Gástrica , Hospedeiro Imunocomprometido , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/diagnóstico , Combinação Trimetoprima e Sulfametoxazol/uso terapêutico , DNA Bacteriano/isolamento & purificação , Humanos , Lactente , Recém-Nascido de Peso Extremamente Baixo ao Nascer/imunologia , Recém-Nascido , Recém-Nascido Prematuro/imunologia , Masculino , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/tratamento farmacológico , Pneumonia por Pneumocystis/imunologia , Pneumonia por Pneumocystis/microbiologia , Reação em Cadeia da Polimerase , Resultado do Tratamento
10.
Parasitol Res ; 118(1): 181-189, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30392033

RESUMO

Pneumocystis jirovecii is an opportunistic fungus occurring in human lungs. The group at highest risk consists of HIV-infected and non-HIV-infected immunosuppressed individuals. In these patients, P. jirovecii infection may lead to Pneumocystis pneumonia; it may, however, persist also in an asymptomatic form. This study aimed to determine the prevalence of P. jirovecii and potential risk factors for infection in a group of renal transplant recipients and to characterize the genetic diversity of this fungus in the studied population. Sputum specimens from 72 patients were tested for presence of P. jirovecii using immunofluorescence microscopy, as well as nested PCR targeting the mtLSU rRNA gene. Genotyping involving analysis of four loci-mtLSU rRNA, CYB, DHPS, and SOD-was used to characterize the diversity of the detected organisms. Pneumocystis DNA was detected in eight (11.11%) patients. It has been shown that low eosinophil count and dual immunosuppressive treatment combining prednisone and calcineurin inhibitors are potential risk factors for colonization. Analysis of genotype distribution showed an association of the wild-type genotype of mtLSU rRNA with lower average age of patients and shorter time after kidney transplantation. Furthermore, CYB 2 genotype was detected only in patients with the ongoing prophylaxis regimen. In conclusion, renal transplant recipients are at risk of Pneumocystis colonization even a long time after transplantation. The present preliminary study identifies specific polymorphisms that appear to be correlated with certain patient characteristics and highlights the need for deeper investigation of these associations in renal transplant recipients.


Assuntos
Transplante de Rim/efeitos adversos , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , Complicações Pós-Operatórias/microbiologia , Adulto , Idoso , Feminino , Variação Genética , Genótipo , Humanos , Hospedeiro Imunocomprometido , Pulmão/microbiologia , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/classificação , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/etiologia , Pneumonia por Pneumocystis/imunologia , Reação em Cadeia da Polimerase , Complicações Pós-Operatórias/imunologia , Prevalência , Transplantados/estatística & dados numéricos , Adulto Jovem
11.
Ann Lab Med ; 39(2): 176-182, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30430780

RESUMO

BACKGROUND: Real-time PCR is more sensitive than microscopic examination for detecting Pneumocystis jirovecii. We compared the performance of two assays for detecting P. jirovecii DNA: the RealStar Pneumocystis jirovecii PCR Kit 1.0 CE (Altona Diagnostics, Hamburg, Germany) and the AmpliSens Pneumocystis jirovecii (carinii)-FRT PCR kit (InterLabService Ltd., Moscow, Russia). METHODS: We used 159 samples from the lower respiratory tract (112 bronchoalveolar lavage [BAL] fluid, 37 sputum, and 10 endotracheal aspirate [ETA] samples) of non-HIV immunocompromised patients. Nested PCR and sequencing were used to resolve discordant results. The performance of the two assays was evaluated according to clinical categories (clinical Pneumocystis pneumonia [PCP], possible PCP, or unlikely PCP) based on clinical and radiological observations. RESULTS: The positive and negative percent agreement values were 100% (95% confidence interval [CI], 85.4-100%) and 96.6% (95% CI, 90.9-98.9%), respectively, and kappa was 0.92 (95% CI, 0.84-0.99). P. jirovecii DNA load was significantly higher in the clinical PCP group than in the other groups (P<0.05). When stratified by sample type, the positive rate for BAL fluids from the clinical PCP group was 100% using either assay, whereas the positive rate for sputum/ETA samples was only 20%. CONCLUSIONS: The two assays showed similar diagnostic performance and detected low P. jirovecii burden in BAL fluids. Both assays may be useful as routine methods for detecting P. jirovecii DNA in a clinical laboratory setting, though their results should be interpreted considering sample type.


Assuntos
Infecções por Pneumocystis/diagnóstico , Pneumocystis carinii/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Líquido da Lavagem Broncoalveolar/microbiologia , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , Erros de Diagnóstico , Humanos , Hospedeiro Imunocomprometido , Infecções por Pneumocystis/microbiologia , Pneumocystis carinii/isolamento & purificação , Kit de Reagentes para Diagnóstico , Doenças Respiratórias/diagnóstico , Doenças Respiratórias/microbiologia , Escarro/microbiologia
12.
BMC Infect Dis ; 19(1): 1092, 2019 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-31888503

RESUMO

BACKGROUND: Varicella-zoster virus (VZV) causes herpes zoster. Pneumocystis jirovecii (PJ) also causes pneumonia in immunocompromised hosts. Although both cause opportunistic infections, it is rare to have a co-infection in a non-human immunodeficiency virus carrier. CASE PRESENTATION: An 84-year-old woman with hemolytic anemia referred because of acute respiratory failure. She had received prednisolone without PJ pneumonia prevention. She developed dyspnea and desaturation while eating, and thus was treated based on a presumptive diagnosis of aspiration pneumonia. Physical examination revealed a vesicular rash on the left side of her neck suggesting herpes zoster infection. Polymerase chain reaction of her sputum for PJ and VZV was positive, which confirmed a diagnosis of pneumonia due to PJ and VZV co-infection. Despite acyclovir and sulfamethoxazole and trimethoprim administration, she died on hospital day 19. CONCLUSIONS: Clinicians should suspect PJP when patients on systemic corticosteroids develop pneumonia and they have not received prophylactic treatment for PJP in non-HIV carriers. When such patients have a VZV rash, clinicians should aggressively seek signs of opportunistic infections. Our case hereby highlights the importance of recognizing the possibility of a VZV and PJ co-infection.


Assuntos
Coinfecção/microbiologia , Coinfecção/virologia , Herpes Zoster/diagnóstico , Herpesvirus Humano 3/genética , Hospedeiro Imunocomprometido/imunologia , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/diagnóstico , Infecção pelo Vírus da Varicela-Zoster/diagnóstico , Aciclovir/administração & dosagem , Aciclovir/uso terapêutico , Idoso de 80 Anos ou mais , Anti-Infecciosos Urinários/administração & dosagem , Anti-Infecciosos Urinários/uso terapêutico , Antivirais/administração & dosagem , Antivirais/uso terapêutico , Coinfecção/tratamento farmacológico , Evolução Fatal , Feminino , Herpes Zoster/tratamento farmacológico , Herpesvirus Humano 3/isolamento & purificação , Humanos , Infecções Oportunistas/tratamento farmacológico , Infecções Oportunistas/microbiologia , Infecções Oportunistas/virologia , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/tratamento farmacológico , Reação em Cadeia da Polimerase , Escarro/microbiologia , Escarro/virologia , Sulfametoxazol/administração & dosagem , Sulfametoxazol/uso terapêutico , Trimetoprima
13.
PLoS One ; 13(10): e0206231, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30359436

RESUMO

BACKGROUND: Pneumocystis jirovecii pneumonia (PCP) is often fatal in human immunodeficiency (HIV)-negative patients and typically presents with respiratory insufficiency. Predicting treatment failure is challenging. This study aimed to identify prognostic factors and examine PCP polymerase chain reaction (PCR)-negative conversion in non-HIV PCP patients with respiratory failure. METHOD: We retrospectively enrolled 81 non-HIV patients diagnosed with and treated for PCP with respiratory failure in the intensive care unit at a tertiary hospital over a 3-year period. PCP was diagnosed via nested PCR-mediated detection of Pneumocystis jirovecii in induced sputum samples, endotracheal aspirates, and bronchoalveolar lavage fluids. PCP PCR was performed weekly to check for negative conversion. RESULTS: The overall survival rate was 35.8%. Seventy-four patients (91.3%) required mechanical ventilation, and 6 (7.4%) required high-flow nasal oxygen treatment. The PCP PCR-negative conversion rate was 70.5% (survivors, 97%; non-survivors, 63.5%); the median time to conversion was 10 (7.0-14.0) days. On univariate analysis, the APACHE II score (p < 0.001), renal failure requiring renal replacement therapy (p = 0.04), PCP PCR-negative conversion (p = 0.003), and the PaO2/FiO2 ratio (first 24 hours) (p < 0.001) significantly correlated with mortality. On multivariate analysis, PCP PCR-negative conversion (hazard ratio, 0.433; 95% confidence interval, 0.203-0.928; p = 0.031) and the PaO2/FiO2 ratio (first 24 hours) (hazard ratio, 0.988; 95% confidence interval, 0.983-0.993; p < 0.001) independently predicted prognosis. CONCLUSIONS: Determination of PCP PCR-negative conversion and PaO2/FiO2 ratios may help physicians predict treatment failure and mortality in non-HIV PCP patients with respiratory failure.


Assuntos
Pneumocystis carinii/genética , Pneumonia por Pneumocystis/complicações , Pneumonia por Pneumocystis/diagnóstico , Pneumonia por Pneumocystis/microbiologia , Síndrome do Desconforto Respiratório do Adulto/complicações , Síndrome do Desconforto Respiratório do Adulto/diagnóstico , Idoso , Feminino , Conversão Gênica , HIV , Soronegatividade para HIV/fisiologia , Humanos , Hospedeiro Imunocomprometido , Unidades de Terapia Intensiva/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/mortalidade , Reação em Cadeia da Polimerase , Prognóstico , Síndrome do Desconforto Respiratório do Adulto/microbiologia , Síndrome do Desconforto Respiratório do Adulto/mortalidade , Estudos Retrospectivos , Análise de Sobrevida
14.
Clin Respir J ; 12(11): 2590-2597, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30244544

RESUMO

INTRODUCTION: Impact of Cytomegalovirus (CMV) co-infection pneumonia in non-HIV patients with Pneumocystis jirovecii pneumonia (PCP) is unclear. OBJECTIVES: The aim of our study was to determine whether CMV co-infection is associated with an increased risk of mortality. METHODS: Our study was conducted at Ege University Hospital, Turkey. We used molecular assays to diagnose Pneumocystis jirovecii in respiratory samples, and CMV in both respiratory and blood samples. We compared morbidity and mortality stratified by CMV co-infection status. RESULTS: Between 2009 and 2015, 43 patients (mean age: 56.7 ± 15.3 years) were diagnosed with PCP. Only 3 of 43 patients had received PCP prophylaxis. We microbiologically confirmed CMV co-infection in 28 of 43 (65.1%) patients. Acute respiratory distress syndrome (ARDS) and requirement of mechanical ventilation were more common in the CMV co-infection group (P = .019 and P = .031 respectively), and duration of intensive care unit was also longer (P = .006). In univariate analyses, mortality at 30 days was higher in the CMV co-infection group as compared to the group with PCP alone (78.6% and 46.7% respectively; P = .046). In multivariate analyses, mortality was independently associated only with the presence of ARDS [OR: 6.22 95% CI 1.3-29.32] and the association with CMV co-infection was no longer significant [OR: 2.6 95% CI 0.49-13.72, P = .257]. CONCLUSION: The risk of mortality appears to be increased in the setting of CMV and PCP co-infection in HIV-uninfected immunocompromised patients. PCP prophylaxis use was lower than expected, suggesting low physician awareness of the risks of PCP in this population.


Assuntos
Coinfecção/complicações , Infecções por Citomegalovirus/epidemiologia , Hospedeiro Imunocomprometido/imunologia , Pneumonia por Pneumocystis/epidemiologia , Adulto , Idoso , Conscientização , Coinfecção/mortalidade , Coinfecção/prevenção & controle , Citomegalovirus/genética , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/mortalidade , Infecções por Citomegalovirus/virologia , Feminino , HIV/classificação , Humanos , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/genética , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , Pneumonia por Pneumocystis/mortalidade , Respiração Artificial/métodos , Síndrome do Desconforto Respiratório do Adulto , Estudos Retrospectivos , Turquia/epidemiologia , Carga Viral
15.
Artigo em Inglês | MEDLINE | ID: mdl-30249686

RESUMO

The echinocandin caspofungin inhibits the catalytic subunit Gsc1 of the enzymatic complex synthesizing 1,3-ß-glucan, an essential compound of the fungal wall. Studies with rodents showed that caspofungin is effective against Pneumocystis asci. However, its efficacy against asci of Pneumocystis jirovecii, the species infecting exclusively humans, remains controversial. The aim of this study was to assess the sensitivity to caspofungin of the P. jirovecii Gsc1 subunit, as well as of those of Pneumocystis carinii and Pneumocystis murina infecting, respectively, rats and mice. In the absence of an established in vitro culture method for Pneumocystis species, we used functional complementation of the Saccharomyces cerevisiae gsc1 deletant. In the fungal pathogen Candida albicans, mutations leading to amino acid substitutions in Gsc1 confer resistance to caspofungin. We introduced the corresponding mutations into the Pneumocystis gsc1 genes using site-directed mutagenesis. In spot dilution tests, the sensitivity to caspofungin of the complemented strains decreased with the number of mutations introduced, suggesting that the wild-type enzymes are sensitive. The MICs of caspofungin determined by Etest and YeastOne for strains complemented with Pneumocystis enzymes (respectively, 0.125 and 0.12 µg/ml) were identical to those upon complementation with the enzyme of C. albicans, for which caspofungin presents low MICs. However, they were lower than the MICs upon complementation with the enzyme of the resistant species Candida parapsilosis (0.19 and 0.25 µg/ml). Sensitivity levels of Gsc1 enzymes of the three Pneumocystis species were similar. Our results suggest that P. jirovecii is sensitive to caspofungin during infections, as are P. carinii and P. murina.


Assuntos
Antifúngicos/farmacologia , Caspofungina/farmacologia , Glucosiltransferases/genética , Pneumocystis carinii/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Teste de Complementação Genética , Glucosiltransferases/metabolismo , Testes de Sensibilidade Microbiana , Mutagênese Sítio-Dirigida , Infecções por Pneumocystis/tratamento farmacológico , Infecções por Pneumocystis/microbiologia , Pneumocystis carinii/efeitos dos fármacos , Subunidades Proteicas/genética , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/genética
16.
Parasitol Res ; 117(10): 3103-3108, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30027382

RESUMO

Sulfonamide group drugs and their antimetabolite combinations are the most preferred drugs in the treatment and prophylaxis of Pneumocystis jirovecii pneumonia (PCP). Especially with the long-term use of trimethoprim-sulfamethoxazole (TMP-SMX) and dapsone, certain point mutations in the Pneumocystis jirovecii (P. jirovecii) dihydropteroate synthase (DHPS) gene are known to play an important role in the development of resistance. In the present study, we investigated the 165th and 171st nucleotide mutations in the DHPS gene in the P. jirovecii isolates from immunosuppressed and immunocompetent cases. P. jirovecii isolates from the bronchoalveolar lavage fluid (BALF) samples of 31 hospitalized cases and lung tissue samples of 37 autopsy cases were included in the study. For the analysis of wild-type and mutant genotypes, after the touchdown-PCR amplification method, the restriction fragment length polymorphism (RFLP) method was used. In this study, P. jirovecii DHPS gene was amplified in 28 of 68 (41%) of the samples. The RFLP method revealed that all the isolates in which the DHPS gene was amplified were considered as wild-type genotypes. To our knowledge, this present study is the first study in Turkey investigating P. jirovecii DHPS gene mutations associated with the sulfonamide resistance. All the isolates showed a wild-type pattern indicating that the occurrence of P. jirovecii DHPS mutations in Turkey is very low or absent.


Assuntos
Antifúngicos/uso terapêutico , Di-Hidropteroato Sintase/genética , Farmacorresistência Fúngica/genética , Pneumocystis carinii/efeitos dos fármacos , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/tratamento farmacológico , Sulfonamidas/uso terapêutico , Adulto , Idoso , Líquido da Lavagem Broncoalveolar , Pré-Escolar , Genótipo , Humanos , Hospedeiro Imunocomprometido , Lactente , Pulmão/parasitologia , Pessoa de Meia-Idade , Mutação/genética , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/microbiologia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Turquia , Adulto Jovem
19.
Rev Esp Quimioter ; 31(4): 336-343, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29956896

RESUMO

OBJECTIVE: The prevalence of Pneumocystis jirovecii colonization and its role in pulmonary disease remains unclear. PCR methods have shown an improved sensitivity in the detection of this fungus. It has been suggested that the PCR results be combined with another test such as IFA to create a diagnostic algorithm. METHODS: A multiplex nested-PCR procedure with a 16S rRNA gene as the internal amplification control was evaluated to determine the role of P. jirovecii in pulmonary disease. RESULTS: A 20% of the 199 bronchoalveolar lavage samples were PCR-positive, 13.5% samples were PCR-inhibited, and the rate of Pneumocystis-colonisation was 6.4%. The sensitivity, specificity, positive predictive value and negative predictive value of the nested-PCR were 100%, 93%, 70% and 100%, respectively. The sensitivity of the nested-PCR was higher than the current "gold standard" immunofluorescence assay (IFA) (p< 0.0001). PCR-negative and PCR-positive patients did not show any clinical or radiological differences in the medical variables studied. CONCLUSIONS: PCR could help the diagnosis of Pneumocystis pulmonary disease given the high negative predictive value of the technique. P. jirovecii DNA can frequently be detected in healthy population, so the analysis of the patient medical history is critical to make the correct clinical decision.


Assuntos
Pneumopatias/complicações , Pneumocystis carinii , Pneumonia por Pneumocystis/complicações , Pneumonia por Pneumocystis/diagnóstico , Reação em Cadeia da Polimerase/métodos , Idoso , Idoso de 80 Anos ou mais , Algoritmos , Líquido da Lavagem Broncoalveolar/microbiologia , DNA Bacteriano/genética , Feminino , Humanos , Pneumopatias/tratamento farmacológico , Pneumopatias/microbiologia , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/genética , Valor Preditivo dos Testes , RNA Bacteriano/genética , RNA Ribossômico 16S/genética
20.
Clin Infect Dis ; 67(6): 913-919, 2018 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-29514207

RESUMO

Background: Although trimethoprim-sulfamethoxazole is the more efficient drug for prophylactic and curative treatment of pneumocystosis, atovaquone is considered a second-line prophylactic treatment in immunocompromised patients. Variations in atovaquone absorption and mutant fungi selection after atovaquone exposure have been associated with atovaquone prophylactic failure. We report here a Pneumocystis jirovecii cytochrome b (cyt b) mutation (A144V) associated with such prophylactic failure during a pneumocystosis outbreak among heart transplant recipients. Methods: Analyses of clinical data, serum drug dosage, and molecular modeling of the P. jirovecii Rieske-cyt b complex were performed to investigate these prophylactic failures. Results: The cyt b A144V mutation was detected in all infected, heart transplant recipient patients exposed to atovaquone prophylaxis but in none of 11 other immunocompromised, infected control patients not treated with atovaquone. Serum atovaquone concentrations associated with these prophylactic failures were similar than those found in noninfected exposed control patients under a similar prophylactic regimen. Computational modeling of the P. jirovecii Rieske-cyt b complex and in silico mutagenesis indicated that the cyt b A144V mutation might alter the volume of the atovaquone-binding pocket, which could decrease atovaquone binding. Conclusions: These data suggest that the cyt b A144V mutation confers diminished sensitivity to atovaquone, resulting in spread of Pneumocystis pneumonia among heart transplant recipients submitted to atovaquone prophylaxis. Potential selection and interhuman transmission of resistant P. jirovecii strain during atovaquone prophylactic treatment has to be considered and could limit its extended large-scale use in immucompromised patients.


Assuntos
Antifúngicos/farmacologia , Atovaquona/farmacologia , Citocromos b/genética , Transplante de Coração , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/etiologia , Adulto , Idoso , Simulação por Computador , Surtos de Doenças , Feminino , Proteínas Fúngicas/genética , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Modelos Moleculares , Mutação , Pneumocystis carinii/efeitos dos fármacos , Pneumocystis carinii/enzimologia , Transplantados , Falha de Tratamento
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