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1.
J. bras. nefrol ; 39(1): 46-54, Jan.-Mar. 2017. tab, graf
Article in English | LILACS | ID: biblio-841202

ABSTRACT

Abstract Introduction: Cytomegalovirus (CMV) infection is a main viral infection after kidney transplantation. The diagnostic methods currently employed are pp65 antigenemia and nucleic acid amplification by polymerase chain reaction (PCR) and aim at detecting viral replication. Objective: The goal of this study was to evaluate and compare by both methods the incidence of CMV active infection in kidney transplant patients and to establishthe best clinical-laboratory correlation. Methods: Thirty sequential kidney transplant recipients were enrolled in a single center prospective cohort study. Peripheral blood samples were drawn from day 15 until the 6th month after transplantation and tested for CMV replication by pp65 antigenemia and quantitative PCR assays (qPCR). Results: Two hundred forty samples were analyzed and the incidence of active infection was similar by both methods. Time elapsed to the first positive test was almost identical but more samples tested positive by qPCR than by antigenemia in a behavior that was almost evenly distributed overtime. Agreement between tests was observed in 217 samples (90.4%; kappa = 0.529; p < 0.001) and in 25 patients the tests were concordant (83.3%; kappa = 0.667; p < 0.001). The evaluation of the diagnostic parameters for CMV replication revealed higher sensitivity for the qPCR test (82.1%) against antigenemia (59.0%). Quantitative PCR was also slightly more accurate than antigenemia. Conclusion: Our data demonstrate that both methods are suitable and have almost equivalent accuracy for the detection of post-transplant cytomegalovirus replication. The choice for either test must take in consideration the demand, execution capability and cost-effectiveness at each institution.


Resumo Introdução: Citomegalovírus (CMV) é uma importante causa de infecção viral após o transplante renal. Os métodos diagnósticos presentemente utilizados são a antigenemia pp-65 e os métodos que utilizam a amplificação de ácidos nucléicos pela reação em cadeia da polimerase (PCR) e visam à detecção da replicação viral. Objetivo: O objetivo deste estudo foi avaliar e comparar a incidência de infecção ativa por CMV em pacientes transplantados renais pelos dois métodos e estabelecer a melhor correlação clínico-laboratorial. Métodos: Trinta pacientes transplantados renais seqüenciais em um único centro foram incluídos em um estudo de coorte prospectiva. Amostras de sangue periférico foram coletadas a partir do 15º dia até o 6º mês pós-transplante e avaliadas para replicação de CMV por Antigenemia pp-65 e PCR quantitativo (qPCR). Resultados: Foram analisadas 240 amostras e a incidência de infecção ativa foi similar pelos dois métodos. O tempo médio transcorrido desde o transplante até o primeiro teste com resultado positivo foi quase idêntico entretanto mais amostras tiveram resultado positivo por qPCR do que antigenemia, um comportamento que se manteve quase uniforme ao longo do tempo. Concordância entre os testes foi observada em 217 amostras (90,4%; kappa = 0,529; p < 0,001) e em 25 pacientes (83,3%; kappa = 0,667; p < 0,001). A avaliação dos parâmetros diagnósticos para replicação de CMV revelaram maior sensibilidade para qPCR (82,1%) contra antigenemia (59,0%). PCR quantitativo também foi levemente mais preciso do que antigenemia. Conclusão: Nossos dados demonstram que ambos os métodos são adequados e tem precisão quase equivalente para a detecção da replicação do CMV após o transplante renal. A escolha entre um ou outro deve levar em consideração a demanda, capacidade de execução e custo-efetividade em cada instituição.


Subject(s)
Humans , Adolescent , Adult , Middle Aged , Young Adult , Postoperative Complications/diagnosis , Postoperative Complications/virology , Kidney Transplantation , Cytomegalovirus Infections/diagnosis , Postoperative Complications/blood , Reproducibility of Results , Longitudinal Studies , Cytomegalovirus Infections/blood , Hematologic Tests
2.
Clin. biomed. res ; 36(1): 18-22, 2016. tab
Article in English | LILACS | ID: lil-788747

ABSTRACT

Introduction: Tuberculosis (TB) persists as a severe global public health issue. The aim of the present study was to evaluate the performance of an in-house TB PCR (polymerase chain reaction) in sputum. Methods: DNA from sputum specimens were submitted to a nested-PCR protocol for the IS6110 region detection. PCR results were compared to those of the traditional methods for TB diagnosis, i.e., acid-fast bacilli (AFB) smear microscopy and culture. We analyzed sputum samples obtained from 133 patients. Results: A total of 48 (36%) cultures yielded indeterminate results due to contamination. This high contamination rate may be explained by the fact that samples from fibrocystic patients were included in this study. Additionally, other five samples were positive for nontuberculous mycobacteria (NTM). Therefore, it was possible to compare 80 patients for M. tuberculosis detection. We found 14 positive samples: five presented positive results in the three methods (5/14; 35.7%), two were positive in culture and PCR (2/14; 14.3%), one was positive in AFB and PCR (1/14; 7.1%), five were positive only in PCR (5/14; 35.7%) and 1 was positive only in culture (1/14; 7.1%). Thus, positivity rates for each technique were: 7.5% for AFB (6/80), 10% for culture (8/80) and 16.25% for PCR (13/80). Among the 48 patients who had indeterminate results in sputum culture, two samples were positive in PCR. Conclusion: Considering the limitations of the traditional methods, the use of PCR as a molecular technique could be advantageous for TB diagnosis.


Subject(s)
Humans , Polymerase Chain Reaction , Tuberculosis, Pulmonary
3.
Clin. biomed. res ; 34(2): 64-68, 2014. graf, tab
Article in English | LILACS | ID: biblio-997748

ABSTRACT

Infection with hepatitis C virus (HCV) is a global public health issue. The bloodborne nature of HCV transmission poses a substantial risk to healthcare workers, due to occupational exposure to needlestick injuries and blood and other body fluids containing the virus. Undiagnosed HCV infection, including in healthcare workers, represents a growing problem worldwide as the infected population ages, and HCV-related mortality and morbidity is expected to rise substantially over the coming decades. Consequently, diagnostic tests for HCV play an important role in this scenario. The aim of this study was to standardize a one-step RT-PCR assay for detection of HCV. The test demonstrated reproducibility, sensibility (100%), and the limit of detection was set at 100IU/mL. Our study indicates that this assay can be used as a diagnostic tool to follow up healthcare workers after occupational exposure


Subject(s)
Humans , RNA, Viral/blood , Hepatitis C/diagnosis , Hepacivirus/isolation & purification , Untranslated Regions/genetics , RNA, Viral/genetics , Hepatitis C/virology , Hepacivirus/genetics , Viral Load/methods
4.
Clin. biomed. res ; 34(3): 260-265, 2014. ilus, tab
Article in Portuguese | LILACS | ID: biblio-834468

ABSTRACT

Introduction: Polyomaviruses (BKV and JCV) cause infection mainly in immunocompromised adults. A sensitive and specific diagnosis tool is fundamental to demonstrate the BKV and JCV infections. Nowadays many laboratories are using a PCR technique for detecting polyomaviruses genome in clinical samples. In this context, the purpose of this study is to determine the threshold of detection of the nested-PCR for polyomaviruses JC and BK. Methods: Serial dilutions of the samples of BKV and JCV of known concentration (100 copies/mL, 50 copies/mL, 25 copies/mL, 10 copies/mL, 5 copies/mL, and 1 copy/ml) were subjected to the technique of nested-PCR. All dilutions were tested 11 times to determine the minimum detection limit. Results: The minimum detection limit of the nested-PCR for JC and BK viruses was 25 copies/mL. This dilution (25 copies/mL) showed 100% PCR positive reaction. Furthermore, we found that weak positive results were obtained at dilutions of 1,5 and 10 copies/mL in some repetitions. Dilutions of 25, 50, and 100 copies/mL always had very positive results. Conclusions: These values are similar to those reported in other studies, contributing to the indication of this PCR for potential diagnostic purposes.


Introdução: Os poliomavírus (JCV e BKV) causam infecções principalmente em adultos imunocomprometidos. Um diagnóstico sensível e específico é de fundamental importância para os pacientes portadores de JCV e BKV. Atualmente alguns laboratórios têm utilizado a técnica de PCR para a detecção do material genético destes vírus em amostras clínicas. Assim, o objetivo deste estudo é determinar o limite mínimo de detecção da técnica de nested-PCR para os poliomavírus JC e BK. Métodos: Diluições seriadas (100 cópias/mL; 50 cópias/mL; 25 cópias/mL; 10 cópias/mL; 5 cópias/mL e 1 cópia/mL) de controles positivos comerciais de JCV e BKV com concentrações conhecidas foram submetidas à técnica de nested-PCR semi-duplex. Todas as diluições foram testadas 11 vezes para determinação do limite mínimo de detecção. Resultados: O limite mínimo de detecção da reação de nested-PCR para os vírus JC e BK foi de 25 cópias/mL para ambos, com 100% de positividade das diluições testadas na reação de PCR. Ainda, pudemos observar que resultados positivos fracos foram obtidos nas diluições de 1, 5 e 10 cópias/mL em algumas das repetições realizadas. As diluições de 25, 50 e 100 cópias/mL sempre obtiveram resultado rancamente positivo. Conclusões: Estes valores são semelhantes aos relatados em outros estudos, contribuindo para a indicação desta reação de PCR para potenciais fins diagnósticos.


Subject(s)
Humans , BK Virus , Polyomavirus Infections/diagnosis , JC Virus , Limit of Detection , Polymerase Chain Reaction , Immunosuppression Therapy , Specimen Handling/standards
5.
Braz. j. infect. dis ; 15(4): 323-327, July-Aug. 2011. ilus, tab
Article in English | LILACS | ID: lil-595672

ABSTRACT

Streptococcus agalactiae or group B Streptococcus (GBS) is one of the most important causal agents of serious neonatal infections. Numerous assays have been evaluated for GBS screening in order to validate a fast and efficient method. The aim of this study was to compare the culture technique (established as the gold standard) with the molecular method of polymerase chain reaction (PCR) with specific primers (atr gene). Two hundred and sixty-three samples were analyzed. Vaginal samples were collected, according to the Centers for Disease Control and Prevention (CDC) recommendations, from women over 35 weeks of pregnancy at Hospital de Clínicas de Porto Alegre (HCPA). Two different extraction methods were tested in all samples collected. PCR technique yielded 71 (26.99 percent) positive results. Sensitivity and specificity for PCR were 100 percent and 86.88 percent, respectively. PCR demonstrated a shorter turnaround time than the culture. The molecular methodology proved to be a useful screening for GBS, allowing effective treatment to be initiated in shorter time to prevent newborn infection.


Subject(s)
Female , Humans , Pregnancy , Culture Media , Polymerase Chain Reaction , Pregnancy Complications, Infectious/diagnosis , Streptococcal Infections/diagnosis , Streptococcus agalactiae/genetics , DNA Primers/analysis , DNA, Bacterial/analysis , Predictive Value of Tests , Pregnancy Complications, Infectious/microbiology , Reagent Kits, Diagnostic , Rectum/microbiology , Sensitivity and Specificity , Streptococcal Infections/microbiology , Streptococcus agalactiae/isolation & purification , Vagina/microbiology
6.
Rev. Soc. Bras. Med. Trop ; 44(3): 286-289, May-June 2011. ilus, tab
Article in English | LILACS | ID: lil-593353

ABSTRACT

INTRODUCTION: Human cytomegalovirus (HCMV) is often reactive in latently infected immunosuppressed patients. Accordingly, HCMV remains one of the most common infections following solid organ and hemopoietic stem cell transplantations, resulting in significant morbidity, graft loss and occasional mortality. The early diagnosis of HCMV disease is important in immunosuppressed patients, since in these individuals, preemptive treatment is useful. The objective of this study was to compare the performance of the in-house qualitative polymerase chain reaction (PCR) and pp65 antigenemia to HCMV infection in immunosuppressed patients in the Hospital de Clínicas of Porto Alegre (HCPA). METHODS: A total of 216 blood samples collected between August 2006 and January 2007 were investigated. RESULTS: Among the samples analyzed, 81 (37.5 percent) were HCMV-positive by PCR, while 48 (22.2 percent) were positive for antigenemia. Considering antigenemia as the gold standard, sensitivity, specificity, positive predictive values and negative predictive values for PCR were 87.5 percent, 76.8 percent, 51.8 percent and 95.5 percent respectively. CONCLUSIONS: These results demonstrated that qualitative PCR has high sensitivity and negative predictive value (NPV). Consequently PCR is especially indicated for the initial diagnosis of HCMV infection. In the case of preemptive treatment strategy, identification of patients at high-risk for HCMV disease is fundamental and PCR can be useful tool.


INTRODUÇÃO: O citomegalovírus humano (HCMV), causador de infecção latente, reativa com frequência em pacientes imunossuprimidos. Portanto, o HCMV permanece uma das infecções mais comuns após transplantes de órgãos sólidos e de células hematopoiéticas resultando em significativa morbidade, perda do enxerto e ocasional mortalidade. Assim, o diagnóstico precoce para uma terapia preventiva é de grande importância. Este estudo visa comparar o desempenho dos métodos PCR qualitativo in-house e antigenemia pp65 para o diagnóstico de infecção por CMV em pacientes imunossuprimidos do Hospital de Clínicas de Porto Alegre. MÉTODOS: O estudo foi realizado em 216 amostras de sangue total (EDTA) coletadas de 85 pacientes, entre agosto de 2006 e janeiro de 2007. RESULTADOS: Dentre as 216 amostras analisadas, 81 (37,5 por cento) amostras apresentaram resultados positivos na PCR, enquanto 48 (22,2 por cento) apresentaram resultados positivos na antigenemia. A sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo para a PCR, considerando antigenemia como padrão foram 87,5 por cento, 76,8 por cento, 51,8 por cento e 95,5 por cento, respectivamente. CONCLUSÕES: Estes resultados demonstraram que a PCR tem alta sensibilidade e valor preditivo negativo. Consequentemente PCR é especialmente indicada para o diagnóstico inicial de infecção por HCMV. No caso da estratégia de terapia preventiva, a identificação de pacientes com alto risco para a doença por HCMV é fundamental e a PCR pode ser uma ferramenta útil.


Subject(s)
Humans , Antigens, Viral/analysis , Cytomegalovirus Infections/diagnosis , Cytomegalovirus/genetics , Cytomegalovirus/immunology , DNA, Viral/analysis , Immunocompromised Host/immunology , Cytomegalovirus Infections/immunology , Predictive Value of Tests , Phosphoproteins/immunology , Polymerase Chain Reaction/methods , Reproducibility of Results , Sensitivity and Specificity , Viral Matrix Proteins/immunology
7.
Rev. HCPA & Fac. Med. Univ. Fed. Rio Gd. do Sul ; 31(2): 138-144, 2011. ilus, tab
Article in English | LILACS | ID: biblio-834406

ABSTRACT

Background: Burkholderia complex (Bcc) infections in cystic fibrosis (CF) patients are associated with decline in lung function and reduced survival. The potential transmissibility of Bcc among CF patients has been reported, indicating that strict segregation of CF patients with Bcc is crucial. Aims: To standardize the PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) assay in order to identify Bcc species and to establish the prevalence of Bcc species and their susceptibility profile among CF patients seen at the Hospital de Clínicas de Porto Alegre (HCPA). Methods: The classification of the clinical isolates recovered from respiratory tract specimens of CF patients as Bcc was achieved using the API-20NE® phenotypic commercial system. The identification of the Bcc species was performed using PCR-RFLP. The antimicrobial disc diffusion susceptibility testing was performed according to the CLSI (2006). Results: API-20NE® was able to identify Bcc isolates (244 specimens), such as B. cepacia, indicating that it was not able to distinguish among the Bcc species. The PCR-RFLP molecular method discriminated the eight reference Bcc species, thus validating the method for clinical isolates. Bcc prevalence determined by PCR-RFLP was 10.6% (26/244). The molecular analysis identified B. cenocepacia in 53.8% (14/26) of infected patients, B. multivorans in 15.4% (4/26), and B. vietnamiensis and B. ambifaria in 7.7% (2/26). The antibiotic resistance profile was variable among Bcc species. Conclusions: The PCR-RFLP method was validated for the identification of Bcc species. B. cenocepacia proved to be the most prevalent species among the CF patients seen at the HCPA.


Introdução: Infecções por bactérias do complexo Burkholderia cepacia (CBC) em pacientes com fibrose cística (FC) estão associadas a declínio da função pulmonar e diminuição da sobrevida. O potencial de transmissibilidade de CBC entre pacientes com FC é uma realidade, tornando-se importante a estrita segregação dos pacientes infectados.Objetivos: Padronizar a técnica de PCR-RFLP (reação em cadeia da polimerase seguida de clivagem com enzimas derestrição) para diferenciação das espécies de CBC e estabelecer a prevalência dessas espécies e seus perfis de sensibilidade em pacientes com FC atendidos no Hospital de Clínicas de Porto Alegre (HCPA). Métodos: A identificação dos isolados clínicos do trato respiratório de pacientes com FC como CBC foi feita pelo sistema deidentificação fenotípica comercial API-20NE®. A diferenciação das espécies de CBC foi realizada por PCR-RFLP, e o teste de suscetibilidade aos antimicrobianos por disco-difusão foi realizado de acordo com o CLSI (2006).Resultados: O sistema API-20NE® identificou todos os isolados do CBC (244 amostras) como B. cepacia, indicando claramente que não distingue as espécies do complexo. O método molecular de PCR-RFLP discriminou as oito espécies de referência de CBC, validando o método para isolados clínicos. A prevalência de CBC por PCR-RFLP foi de 10,6% (26/244).A análise molecular apontou B. cenocepacia colonizando em 53,8% (14/26) dos pacientes infectados, B. multivorans em 15,4% (4/26) e B. vietnamiensis e B. ambifaria em 7,7% (2/26). O perfil de resistência entre as espécies de CBC para os antibióticos testados foi variado. Conclusão: Foi validada a aplicação do método molecular PCR-RFLP para identificar espécies de CBC, e B. cenocepaciafoi a espécie mais prevalente entre os pacientes fibrocísticos atendidos no HCPA.


Subject(s)
Humans , Burkholderia cepacia complex/genetics , Cystic Fibrosis , Polymorphism, Restriction Fragment Length , Polymerase Chain Reaction/methods , Burkholderia Infections/diagnosis , Burkholderia Infections/microbiology
8.
Braz. j. infect. dis ; 14(6): 564-568, Nov.-Dec. 2010. ilus, graf, tab
Article in English | LILACS | ID: lil-578431

ABSTRACT

INTRODUCTION: Resistance to macrolides, lincosamides and streptogramins B (MLS B antibiotics) in staphylococci may be due to modification in ribosomal target methylase encoded by erm genes. The expression of MLS B resistance lead to three phenotypes, namely constitutive resistance (cMLS B), inducible resistance (iMLS B), and resistance only to macrolides and streptogramins B (MS B). The iMLS B resistance is the most difficult to detect in the clinical laboratory. OBJECTIVE: This study investigated the expression of MLS B resistance and the prevalence of the erm genes among 152 clinical isolates of Staphylococcus aureus and coagulase-negative Staphylococcus (CNS) from Hospital de Clínicas de Porto Alegre. METHODS: Primary MLS B resistance was detected by the disk diffusion method. Isolates with iMLS B phenotype were tested by double-disk induction method. All isolates were tested by a genotypic assay, PCR with specific primers. RESULTS: A total of 46.7 percent of staphylococci were positive for cMLS B; 3.3 percent for iMLS B and 3.3 percent for MS B. One or more erm genes were present in 50.1 percent of isolates. The gene ermA was detected in 49 isolates, ermC in 29 and ermB in 3. CONCLUSION: The prevalence of the ermA, ermB and ermC genes were 29.6 percent, 17.1 percent and 0.66 percent respectively, and constitutive resistance was the most frequent as compared to the other two phenotypes.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial/genetics , Genes, Bacterial/genetics , Macrolides/pharmacology , Staphylococcus/drug effects , Staphylococcus/genetics , Bacterial Proteins/genetics , Coagulase/metabolism , Disk Diffusion Antimicrobial Tests , Genotype , Genes, Bacterial/drug effects , Phenotype , Polymerase Chain Reaction , Staphylococcus/enzymology
9.
Rev. Soc. Bras. Med. Trop ; 43(4): 377-381, jul.-ago. 2010. graf, tab
Article in English | LILACS | ID: lil-556000

ABSTRACT

INTRODUCTION: Bacterial colonization of the lungs is the main cause of morbidity in cystic fibrosis (CF). Pathogens such as Staphylococcus aureus are very well adapted to the pulmonary environment and may persist for years in the same patient. Genetic determinants of these bacteria, such as the presence of SCCmec have recently emerged as a problem in this population of patients. METHODS: Staphylococcus aureus isolates obtained from different clinical materials coming from CF and non-CF patients attended at a cystic fibrosis reference hospital were compared according to SCCmec type and antibiotic susceptibility profile. RESULTS: Three hundred and sixty-four single-patient Staphylococcus aureus isolates were collected, of which 164 (45 percent) were from CF patients. Among the latter, 57/164 (44.5 percent) were MRSA, and among the non-CF patients, 89/200 (35 percent) were MRSA. Associated pathogens were found in 38 CF patients. All 57 MRSA from CF patients harbored the multiresistant cassette type III. In contrast, 31/89 MRSA from non-CF patients harbored SCCmec type I (35 percent) and 44/89 harbored type III (49 percent). The antibiotic susceptibility pattern was similar between CF and non-CF patients. CONCLUSIONS: The high prevalence of multiresistant SCCmec type III among CF patients compared with non-CF patients in our institution may make it difficult to control disease progression through antibiotic therapy for promoting the survival of this kind of patient.


INTRODUÇÃO: Colonização pulmonar bacteriana é a principal causa de morbidade em fibrose cística (FC). Patógenos como Staphylococcus aureus são muito bem adaptados ao ambiente pulmonar e podem persistir por anos no mesmo paciente. Determinantes genéticos desta bactéria, como presença de SCCmec emergiram recentemente como um problema nesta população de pacientes. MÉTODOS: Foram comparados isolados de Staphylococcus aureus obtidos de diferentes materiais clínicos, de pacientes com e sem FC atendidos em um hospital de referência em tratamento de fibrocísticos de acordo com o tipo de SCCmec e o perfil de susceptibilidade aos antimicrobianos. RESULTADOS: Foram coletados 364 Staphylococcus aureus, um isolado por paciente, sendo 164 (45 por cento) de pacientes com FC. Entre estes pacientes, 57/164 (44,5 por cento) eram MRSA, e entre pacientes não fibrocísticos, MRSA compreendiam 89/200 (35 por cento). Foram encontrados outros patógenos, associados ao Staphylococcus aureus, em 38 pacientes com FC. Todos 57 MRSA de pacientes com FC possuíam o cassete de multiresistência tipo III. Por outro lado, 31/89 MRSA de pacientes não fibrocísticos possuíam SCCmec tipo I (35 por cento) e 44/89 possuíam tipo III (49 por cento). O perfil de susceptibilidade aos antimicrobianos foi similar entre pacientes com e sem FC. CONCLUSÕES: A alta prevalência do SCCmec de multiresistência tipo III entre pacientes com FC comparado com pacientes sem FC em nossa instituição pode dificultar o controle da progressão da doença, feito através da antibioticoterapia, e que promove a sobrevivência deste tipo de paciente.


Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Cystic Fibrosis/microbiology , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/epidemiology , Brazil/epidemiology , Microbial Sensitivity Tests , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/genetics , Prevalence , Staphylococcal Infections/microbiology
10.
Article in English | LILACS | ID: lil-552737

ABSTRACT

Pulmonary toxoplasmosis is a challenging diagnosis in immunosuppressed patients with nonspecific clinical picture and radiologic findings. We present a case of pneumonia due to Toxoplasma gondii diagnosed by polymerase chain reaction (PCR) in the bronchoalveolar lavage (BAL) fluid of a patient with acquired immunodeficiency syndrome (AIDS). Coinfection with Pneumocystis jirovecii was found in the same specimen. Direct examination and culture for bacteria, mycobacteria and other fungus were negative. Despite the intensive management, respiratory compromise evolved rapidly, with the need for ventilatory support. Acute respiratory distress syndrome developed, and the patient died of multiple organ failure. This case illustrates that a high index of suspicion is necessary for diagnosis of pulmonary toxoplasmosis, a potentially fatal condition. Due to high diagnostic performance, PCR in BAL fluid should be included in the evaluation of immunosuppressed patients with nonspecific pulmonary diseases.


O diagnóstico de toxoplasmose pulmonar em pacientes imunossuprimidos é difícil, devido ao quadro clínico e aos achados radiológicos inespecíficos. Neste artigo, relatamos o caso de uma paciente com síndrome da imunodeficiência adquirida (SIDA), que apresentou pneumonia por Toxoplasma gondii diagnosticada através de reação em cadeia da polimerase (PCR) no lavado bronco-alveolar (LBA). A paciente apresentava co-infecção com Pneumocystis jirovecii. Os demais exames microbiológicos, como bacterioscópico, cultural para bactérias, micobactérias e fungos, foram negativos. Apesar do manejo intensivo, a paciente evoluiu com síndrome do desconforto respiratório agudo e óbito por falência múltipla dos órgãos. Este caso demonstra que um alto índice de suspeita clínica é necessário para o diagnóstico de pneumonia por Toxoplasma gondii. Devido ao seu desempenho diagnóstico, o PCR para Toxoplasma gondii no LBA deve ser incluído na avaliação de pacientes imunossuprimidos com quadros pulmonares inespecíficos.


Subject(s)
Humans , Female , Middle Aged , Polymerase Chain Reaction , Pneumocystis carinii/pathogenicity , Acquired Immunodeficiency Syndrome/complications , Acquired Immunodeficiency Syndrome/diagnosis , Acquired Immunodeficiency Syndrome/epidemiology , Acquired Immunodeficiency Syndrome/mortality , Acquired Immunodeficiency Syndrome/pathology , Toxoplasmosis/diagnosis , Toxoplasmosis/mortality , Toxoplasmosis/pathology , Bronchoalveolar Lavage/instrumentation , Bronchoalveolar Lavage/methods , Bronchoalveolar Lavage , Pneumonia/complications , Pneumonia/diagnosis , Pneumonia/mortality , Pneumonia/pathology , Pneumonia/prevention & control
11.
Article in Portuguese | LILACS | ID: lil-566995

ABSTRACT

Mundialmente, a hepatite pelo vírus B (HBV) é considerada um dos maiores problemas de saúde pública, apesar da vacinação. A Organização Mundial da Saúde (OMS) estima que mais de 2 bilhões de pessoas estejam infectadas pelo HBV. O Brasil é classificado como área de incidência intermediária pela OMS. No entanto, estudos de prevalência detectaram diferenças de índices de infecção nas regiões geográficas: 8% na região Amazônica, 2,5% nas regiões Centro-Oeste e Nordeste, 2% na Sudeste e 1% na região Sul. Um diagnóstico sensível e específico é de fundamental importância para os pacientes portadores do HBV. O objetivo deste estudo foi determinar o limite mínimo de detecção da técnica de PCR “nested” “in house” para o HBV. Diluições seriadas de uma amostra quantificada de HBV (1000 cópias/mL; 750 cópias/mL; 500 cópias/mL; 250 cópias/mL) foram submetidas à técnica de PCR “nested”. O alvo da amplificação por PCR foi a região do core e pré-core do vírus. Para extração dos ácidos nucléicos da amostra foi empregado o kit comercial QIAmp. O limite mínimo de detecção encontrado foi de 500 cópias/mL ou 10 cópias por reação de PCR.


All over the world, the hepatitis B virus (HBV) is considered one of the major problems of public health, despite vaccination. World Health Organization (WHO) estimates that more than 2 billions of persons are infected by HBV. Brazil is classified as an area of intermediary incidence by WHO. However, prevalence studies have detected differences of infection indexes in geographic regions: 8% in the Amazonian region, 2,5% in middle-west and Northeast, 2% in Southeast and 1% in South. A sensitive and specific diagnosis is very important to the HBV carrier patients. The aim of this study was to determine the minimum limit of detection of the nested PCR in house technique for HBV. Serial dilutions of one quantified sample of HBV (1000 copies/mL; 750 copies/mL; 500 copies/mL; 250 copies/mL) were submitted to a nested PCR. The target of PCR was viral core and pre-core region. Commercial kit, QiAmp, was employed to purify nucleic acids from the sample. The minimum detection limit found was 500 copies/mL or 10 copies per PCR reaction.


Subject(s)
Humans , Cross-Sectional Studies , Hepatitis B/diagnosis , Hepatitis B/epidemiology , Hepatitis B/pathology , Incidence , Polymerase Chain Reaction/methods
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