Chlamydophila pneumoniae infection is not associated with primary biliary cholangitis / La infección por Chlamydophila pneumoniae no está asociada con colangitis biliar primaria

ANTECEDENTES: La colangitis biliar primaria (CBP) es una enfermedad hepática inflamatoria crónica colestásica de causa desconocida. Varios patógenos virales y bacterianos han sido propuestos como factores que podrían gatillar una respuesta inmune por mimetismo molecular, o directamente estar relacionados en la persistencia del daño biliar. Existen reportes controversiales respecto al rol de en la patogenia de CBP. OBJETIVOS: Investigar marcadores de infección de séricos y en hígado de pacientes con CBP. PACIENTES Y MÉTODOS: Veinte pacientes diagnosticados con CBP y 20 pacientes control con otras enfermedades hepáticas crónicas no colestásicas fueron estudiados. Se determinaron anticuerpos séricos anti- (IgG). Se realizó detección inmunohistoquímica de antígenos de en hígado. Se extrajo DNA de hígado para amplificación de la secuencia específica de rRNA 16S de por PCR. Fueron usados controles de amplificación de DNA bacteriano y humano. Los pacientes firmaron consentimiento informado. Se realizó un metaanálisis de la diferencia de riesgo de CBP en pacientes infectados por y en un grupo control. RESULTADOS: Los anticuerpos séricos fueron positivos en 30% de los pacientes con CBP y 50% de los controles (p = NS). Antígenos de no fueron detectados en tejido hepático de pacientes con CBP ni de controles. No se amplificó ADN bacteriano en ninguna de las muestras. El metaanálisis de la diferencia de riesgo mostró gran heterogeneidad de los estudios, por lo que no se realizó una estimación de diferencia de riesgo agrupada. DISCUSIÓN: No encontramos asociación entre infección por y CBP. En la evidencia actual, un estudio presenta resultados a favor de la asociación entre y CBP y tres estudios resultados en contra.,

Primary biliary cholangitis (PBC) is a chronic cholestatic inflammatory liver disease of unknown cause. Several viral and bacterial pathogens have been proposed as factors that could either trigger an immune response by molecular mimicry or directly be involved in the persistence of biliary damage. There are conflicting reports respecting the role of in the pathogenesis of PBC. To investigate markers of infection in serum and liver tissue from patients with PBC. Twenty patients with diagnosis of PBC and 20 control patients with other non-cholestatic chronic liver diseases were studied. Serum anti- antibodies (IgG) were determined. Liver tissue was available for immunohistochemistry detection of antigens. DNA was extracted from liver tissue and a specific sequence of 16S rRNA gene was amplified by CPR. Adequate controls of bacterial and human DNA amplification were used. Informed consent was obtained from patients. A meta-analysis of risk difference of PBC in Chlamydophila pneumoniae infected patients and in the control groupwas performed. Serum antibodies were positive in 30% of patients with PBC and 50% of controls (p = NS). antigens were not detected in liver tissue neither of patients with PBC nor controls. Bacterial DNA did not amplify in any of the samples, despite good amplification of internal and external controls. Risk difference meta-analysis showed high heterogeneity between studies. Therefore, we did not estimate a pooled risk difference. Our results do not support the association between infection and PBC. In the current literature only one study shows an association between and PBC, but other three studies do not support it.

Role of Atypical Pathogens and the Antibiotic Prescription Pattern in Acute Bronchitis: A Multicenter Study in Korea

The role of atypical bacteria and the effect of antibiotic treatments in acute bronchitis are still not clear. This study was conducted at 22 hospitals (17 primary care clinics and 5 university hospitals) in Korea. Outpatients (aged > or = 18 yr) who had an acute illness with a new cough and sputum (< or = 30 days) were enrolled in 2013. Multiplex real-time polymerase chain reaction (RT-PCR) was used to detect five atypical bacteria. A total of 435 patients were diagnosed as having acute bronchitis (vs. probable pneumonia, n = 75), and 1.8% (n = 8) were positive for atypical pathogens (Bordetella pertussis, n = 3; B. parapertussis, n = 0; Mycoplasma pneumoniae, n = 1; Chlamydophila pneumoniae, n = 3; Legionella pneumophila, n = 1). Among clinical symptoms and signs, only post-tussive vomiting was more frequent in patients with atypical pathogens than those without (P = 0.024). In all, 72.2% of the enrolled patients received antibiotic treatment at their first visits, and beta-lactams (29.4%) and quinolones (20.5%) were the most commonly prescribed agents. In conclusion, our study demonstrates that the incidence of atypical pathogens is low in patients with acute bronchitis, and the rate of antibiotic prescriptions is high.

Comparison of Sputum and Nasopharyngeal Swab Specimens for Molecular Diagnosis of Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophila

BACKGROUND: Differentiation of atypical pathogens is important for community-acquired pneumonia (CAP). In this study, we compared sputum and nasopharyngeal swabs (NPS) for use in detection of Mycoplasma pneumoniae (MP), Chlamydophila pneumoniae (CP), and Legionella pneumophila (LP), using Seeplex PneumoBacter ACE Detection Assay (PneumoBacter; Seegene). METHODS: Sputum and NPS specimens were collected from patients in 15 hospitals. DNA was extracted from sputum using QIAamp DNA Stool Mini Kit (Qiagen) and from NPS using easyMAG (bioMerieux). Both types of specimens were evaluated by multiplex PCR using PneumoBacter. To determine the diagnostic performance of this assay, sputum samples were also tested using BD ProbeTec ET Atypical Pneumonia Assay (APA; Becton Dickinson). RESULTS: Among 217 sputum and NPS, 20 (9.2%), 2 (0.9%), and 0 sputum were positive for MP, LP, and CP, respectively, whereas 8 (3.7%) NPS were positive for MP. The sputum APA test yielded 186, 206, and 204 interpretable results for MP, LP, and CP, respectively. Of these, 21 (11.3%) were positive for MP, 2 (1.0%) were positive for LP, and 0 samples were positive for CP. Compared to APA, the sensitivity and specificity of the sputum assay for MP were 95.2% and 100.0%, respectively, whereas for the NPS assay, these were 38.1% and 93.9%. Sputum testing was more sensitive than NPS testing (P=0.002). For LP and CP diagnosis, PneumoBacter and APA tests agreed 100%. CONCLUSIONS: Specimen type is crucial and sputum is preferred over NPS for simultaneous detection of MP, LP, and CP using multiplex PCR in CAP.

Inflamação sistêmica causada pela periodontite crônica em pacientes vítimas de ataque cardíaco isquêmico agudo / Systemic inflammation caused by chronic periodontite in patients victims of acute ischemic heart attack

OBJETIVO: Processos inflamatórios e infecciosos mediados por bactérias em sítios distantes têm sido descritos como fator de risco à doença coronariana isquêmica aguda (DCIA). MÉTODOS: Cento e oitenta e um pacientes com DCIA, com e sem periodontites crônicas, foram incluídos neste estudo. Os pacientes foram admitidos no HC da UNICAMP e estratificados em três grupos: grupo 1 - pacientes com periodontite crônica grave (31 homens e 19 mulheres; média de idade 55,1 ± 11,29 anos); grupo 2 - pacientes com periodontite crônica leve (40 homens e 28 mulheres; média de idade 54,8 ± 10,37 anos); grupo 3 - pacientes desdentados (43 homens e 20 mulheres; média de idade 67,5 ± 8,55 anos). Amostras sanguíneas foram coletadas para mensurar os perfis lipídico, hematológico e glicêmico. Além disso, biópsias de 17 artérias coronárias com aterosclerose e igual número de artérias mamárias internas sem degeneração aterosclerótica no grupo 1 foram investigadas. Para análise estatística utilizou-se a análise de variância (ANOVA) e o teste de Scheffé para comparações múltiplas. RESULTADOS: Triglicérides e LDL estavam elevados no grupo 1 em relação ao grupo 2. O HDL apresentou-se reduzido em 20 por cento dos pacientes do grupo 1, e em 8 por cento nos desdentados. A glicemia estava elevada no grupo 1. DNA de bactérias periodontais foram detectados em 58,8 por cento das artérias coronárias. CONCLUSÕES: Pacientes com DCIA e periodontite crônica grave podem apresentar perfil lipídico alterado, como também microorganismos associados com as periodontites crônicas graves podem permear dentro de vasos coronarianos.

OBJECTIVE: Infectious and inflammatory processes mediated by bacteria in distant sites have been described as a risk factor for acute ischemic heart disease (AIHD). METHODS: One hundred one patients with AIHD with and without chronic periodontitis (CP) were included in this study. Patients were admitted to the HC UNICAMP and stratified into three groups: in group 1, we selected patients with severe chronic periodontitis (31 men and 19 women, mean age 55.1 ± 11.29 years old); the group 2 with mild chronic periodontitis (40 men and 28 women, mean age 54.8 ± 10.37 years old) and group 3 represented by the toothless (43 men and 20 women, mean age 67.5 ± 8.55 years old). Blood samples were collected to measure the lipid profiles, hematological and blood glucose levels. In addition, biopsies of seventeen coronary arteries with atherosclerosis and an equal number of internal mammary arteries without atherosclerotic degeneration in group 1 were investigated. Statistical analysis by analysis of variance (ANOVA) and Scheffé test for multiple comparisons was performed. RESULTS: Triglyceride and LDL levels were elevated in group 1 than in group 2. HDL were reduced by 20 percent in group 1 and remained reduced by 8 percent in toothless. Blood glucose was higher in group 1. DNA of periodontal bacteria was detected in 58.8 percent of the coronary arteries. CONCLUSIONS: Patients with (AIHD) and severe chronic periodontitis may have altered lipid profile, as well as microorganisms associated with CP can permeate into coronary vessels.

Chlamydia pneumoniae and stroke: is there a direct relationship? / Chlamydia pneumoniae e acidente vascular cerebral aterotrombótico: existe relação direta?

OBJECTIVE: To investigate the possible relationship between atherothrombotic stroke and Chlamydia pneumoniae. METHOD: 150 patients with carotid atherothrombosis were enrolled. The casuistic was divided in three groups: ischemic stroke (IS): 65 patients; transient ischemic attack (TIA): 26 patients; and control: 59. The IS or TIA onset was up to 30 days from the beginning of the study. Carotid atheromatoses was diagnosed by Doppler-ultrasonography. Patients with cardioembolic risk or non-atherothrombotic origin were excluded. Comparisons were done between the three groups, and within each group according to the different age sub-groups, to the main arteries affected, and to the atherogenic risk factors. Bacteria detection was done using polimerase chain reaction. RESULTS: Only one patient tested positive for C. pneumoniae belonging to the control group. CONCLUSION: These results do not suggest that C. pneumoniae participated in the onset of IS or TIA or that it has a role in carotid plaque destabilization.

OBJETIVO: Investigar a possível relação entre Chlamydia pneumoniae e acidente vascular cerebral aterotrombótico (AVC). MÉTODO: 150 pacientes com aterotrombose carotídea foram estudados. A casuística foi dividida em 3 grupos: AVC: 65 pacientes; ataque isquêmico transitório (AIT): 26 pacientes e controles: 59. O início do AVC ou AIT era até 30 dias da inclusão no estudo. A ateromatose carotídea foi diagnosticada por ultrassonografia com Doppler. Os pacientes com risco cárdio-embólico ou sem evidência de aterotrombose foram excluídos. Foram estabelecidas comparações entre os 3 grupos e dentro de cada grupo, formado sub-grupos de acordo com diferentes idades, território arterial comprometido e fatores de risco. A detecção da bactéria foi feita por reação de polimerização em cadeia. RESULTADOS: Somente um paciente, pertencente ao grupo controle, teve resultado positivo. CONCLUSÃO: Estes achados não sugerem que a C. pneumoniae participe no desencadeamento do AVC ou AIT ou que tenha papel na desestabilização da placa.

Chlamydia pneumoniae and symptomatic carotid atherosclerotic plaque: a prospective study

OBJECTIVE: To investigate the possible link between symptomatic carotid atherosclerotic plaque and Chlamydia pneumoniae. BACKGROUND: Recently, several studies have demonstrated that there may be a possible link between Chlamydia pneumonia and carotid atherosclerosis, however the real role of Chlamydia pneumoniae is not completely understood. METHOD: This is a prospective study with a total of 52 patients analyzed. All patients had been submitted to endarterectomy, and had suffered thrombotic ischemic stroke or transient ischemic attack up to 60 days prior to the surgery. Every patient presented carotid stenosis over 70 percent. The plaque was removed during the surgery and the laboratory exams were immediately done. Evaluation of Chlamydia pneumoniae DNA was done using polymerase chain reaction (PCR). RESULTS: The PCR analyses of all 52 patients were negative for Chlamydia pneumoniae. CONCLUSION: These initial results do not show a relationship between Chlamydia pneumoniae and symptomatic carotid atherosclerotic plaque.

OBJETIVO: Investigar a possível relação entre placa sintomática de carótidas e Chlamydia pneumoniae. INTRODUÇÃO: Vários estudos têm demonstrado uma possível relação entre Chlamydia pneumonia e aterosclerose carotídea, entretanto o papel definitivo da bactéria não é totalmente conhecido. Há muita especulação: poderia iniciar o processo aterosclerótico, agravá-lo ou desestabilizá-lo. MÉTODO: Estudo prospectivo com um total de 52 pacientes, endarterectomizados e previamente acometidos de acidente vascular cerebral isquêmico ou crise isquêmica transitória, em até 60 dias antes da cirurgia. Todos os pacientes apresentavam estenose carotídea superior a 70 por cento. Os testes laboratoriais foram realizados imediatamente após a endarterectomia. A Chlamydia pneumoniae foi pesquisada através de exame de DNA com reação de polimerização em cadeia (PCR). RESULTADOS: O PCR dos 52 pacientes foram negativos para Chlamydia pneumoniae. CONCLUSÃO: Estes resultados iniciais não mostram relação entre Chlamydia pneumoniae e desestabilização de placa aterosclerótica das carótidas.

Optimization of a polymerase chain reaction (PCR) for increasing its sensitivity to detect Chlamydia pneumoniae specific genome.

Being an intracellular parasite, Chlamydia pneumoniae disseminates to organs outside the respiratory tract and causes chronic diseases in human. Nucleic acid-based method such as polymerase chain reaction (PCR) as diagnostic test has greater sensitivity and specificity than conventional microbiological techniques. The PCR protocol consisting of touchdown technique to detect C. pneumoniae DNA using major outer membrane protein gene (MOMP) was carried out in our laboratory as described in reference paper, but analytical sensitivity reported in it was not reproducible. Hence, the PCR was optimized after modifications in annealing temperature and magnesium ion concentrations. First round PCR profile with annealing at 56 degrees C for 8 cycles followed by 32 cycles with annealing temperature maintained at 54 degrees C and second round profile modified with annealing temperature maintained at 49 degrees C had resulted in 3-fold increase in clinical sensitivity. The present work highlights the importance of optimization of PCR in laboratory settings.

Prevalence and clinical presentations of atypical pathogens infection in community acquired pneumonia in Thailand.

OBJECTIVES: To determine the prevalence of atypical pneumonia and clinical presentations in patients with community acquired pneumonia (CAP). MATERIAL AND METHOD: A prospective multi-centered study was performed in patients aged > or = 2 years with the diagnosis of CAP who were treated at seven governmental hospitals in Bangkok from December 2001 to November 2002. The diagnosis of current infection was based on > or = 4 fold rise in antibody sera or persistently high antibody titers together with the presence of DNA of M. pneumoniae or C. pneumoniae in respiratory secretion or antigen of L. pneumophila in the urine. Clinical presentations were compared between patients with atypical pneumonia and unspecified pneumonia. RESULTS: Of 292 patients, 18.8% had current infection with atypical respiratory pathogens (M. pneumoniae 14.0%, C. pneumoniae 3.4%, L. pneumophila 0.4% and mixed infection 1.0%). Only age at presentation was significantly associated with atypical pneumonia in adults, while absence of dyspnea, lobar consolidation, and age > or = 5 years were significant findings for atypical pneumonia in children. CONCLUSION: The present study confirms the significance of atypical pathogens in adults and children. Moreover lobar consolidation is likely to predict atypical pneumonia in childhood CAP.

Atypical Pathogens as Etiologic Agents in Hospitalized Patients with Community-Acquired Pneumonia in Korea: A Prospective Multi-Center Study

Local epidemiologic data on the etiologies of patients hospitalized with community-acquired pneumonia (CAP) is needed to develop guidelines for clinical practice. This study was conducted prospectively to determine the proportion of atypical bacterial pathogens in adults patients hospitalized with CAP in Korea between October 2001 and December 2002. Microbiological diagnosis was determined by serology for antibodies to Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneu-mophila. Nucleic acid of M. pneumoniae and C. pneumoniae in respiratory samples and Legionella antigen in urine samples were detected. The study population consisted of 126 patients (71 males, 55 females), averaging 54.6 yr (SD+/-17.8), whose paired sera were available. An etiologic diagnosis for atypical pathogens was made in 18 patients (14.3%): C. pneumoniae 9 (7.1%), M. pneumoniae 8 (6.3%), and L. pneumophila 3 patients (2.4%). Streptococcus preumoniae and other typical pathogens were isolated from 36 patients (28.6%). Of 126 patients, 16 (12.7%) were admitted to intensive care unit and atypical pathogens were identified in 5 patients (31.3%). Initial clinical features of patients with pneumonia due to atypical, typical or undetermined pathogens were indistinguishable. We conclude that atypical pathogens should be seriously considered in hospitalized patients with CAP, when initiating empiric treatment in Korea.

Rapid detection of Chlamydia pneumoniae DNA in peripheral blood mononuclear cells of coronary artery disease patients by real-time fluorescence PCR.

Several recent reports including serological, pathological and animal studies have associated Chlamydia pneumoniae with coronary artery disease (CAD). In order to establish whether chronic C. pneumoniae infection is linked to coronary artery disease, clinical intervention trials may be needed. However, to detect eligible patients with persistent infection, a reliable diagnostic marker must be developed for identifying cases and assessing efficacy of antichlamydial therapy. Moreover, the prevalence of circulating C. pneumoniae DNA in CAD patients varied widely from previous reports. A real-time PCR has been established by using HL-1 and HR-1 primer to amplify 437 base pairs product. Confirmation of the product was performed on LightCycler by melting curve analysis of detection probes labeled with LC-Red705. Ninety-five angiographically confirmed CAD patients and 104 normal, healthy volunteers were recruited. The mononuclear cell layer was separated from collected blood and rapid, single step real-time PCR was used to detect C. pneumoniae DNA. C. pneumoniae DNA in peripheral blood mononuclear cells (PBMC) was found in 17 per cent of 95 CAD patients and 1 per cent of 104 normal healthy volunteers (odds ratio 20.86, 95% confidence interval 2.71 - 160.67, p < 0.0001). There was no association between C. pneumoniae DNA in PBMC and serological status. The rapid, real-time PCR showed a clear-cut result between positive and negative cases. PBMC-based real-time PCR may be a useful tool for identifying subjects carrying C. pneumoniae in the circulation or in the vascular wall as well. It will be a specific indicator of current infection and will be used as a marker for assessing the microbiological efficacy of antichlamydial therapy in clinical intervention trials.