Assessing and screening for T-cell epitopes from Mycobacterium tuberculosis RD2 proteins for the diagnosis of active tuberculosis

ABSTRACT The Region of D eletion 2 (RD2) of Mycobacterium tuberculosis encodes reserved antigens that contribute to bacterial virulence. Among these antigens, Rv1983, Rv1986, Rv1987, and Rv1989c have been shown to be immunodominant in infected cattle; however, their diagnostic utility has not been evaluated in humans.In this study, we screened 87 overlapping synthetic peptides encoded by five RD2 proteins for diagnosing tuberculosis epitopes in 50 active tuberculosis (TB) cases, 31 non-tuberculosis patients and 36 healthy individuals. A pool of promising epitopes was then assessed for their diagnostic value in 233 suspected TB patients using a whole blood IFN-γ release assay.Only 10 peptides were recognized by more than 10% of active tuberculosis patients. The IFN-γ release responses to Rv1986-P9, P15, P16, Rv1988-P4, P11, and Rv1987-P11 were significantly higher in the active TB group than in the control groups (p < 0.05). The whole blood IFN-γ release assay based on these epitopes yielded a sensitivity of 51% and a specificity of 85% in diagnosing active tuberculosis, and the corresponding results using the T-SPOT.TB assay were 76% and 75%, respectively.In conclusion, these results suggest that the six epitopes from the RD2 of M. tuberculosis have potential diagnostic value in TB.

The role of environmental tobacco exposure and Helicobacter pylori infection in the risk of chronic tonsillitis in children / O papel da exposição ambiental do tabaco e infecção pelo Helicobacter pylori no risco de amigdalite crônica em crianças

ABSTRACT CONTEXT AND OBJECTIVE: Helicobacter pylori (H. pylori) is a chronic infectious pathogen with high prevalence. This study investigated the interaction between environmental tobacco exposure and H. pylori infection on the incidence of chronic tonsillitis in Chinese children. DESIGN AND SETTING: Cross-sectional study performed in an outpatient clinic in China. METHODS: Pediatric patients with chronic tonsillitis were enrolled. H. pylori infection was determined according to the presence of H. pylori CagA IgG antibodies. Serum cotinine levels and environmental tobacco smoke (ETS) exposure were determined for all participants. RESULTS: There was no significant difference in H. pylori infection between the children with chronic tonsillitis and children free of disease, but there was a significant difference in ETS between the two groups (P = 0.011). We next studied the association between ETS and chronic tonsillitis based on H. pylori infection status. In the patients with H. pylori infection, there was a significant difference in ETS distribution between the chronic tonsillitis and control groups (P = 0.022). Taking the participants without ETS as the reference, multivariate logistic regression analysis showed that those with high ETS had higher susceptibility to chronic tonsillitis (adjusted OR = 2.33; 95% CI: 1.67-3.25; adjusted P < 0.001). However, among those without H. pylori infection, ETS did not predispose towards chronic tonsillitis. CONCLUSION: Our findings suggest that tobacco exposure should be a putative mediator risk factor to chronic tonsillitis among children with H. pylori infection.

RESUMO CONTEXTO E OBJETIVO: Helicobacter pylori (H. pylori) é um patógeno infeccioso crônico com alta prevalência. Este estudo investigou a interação entre exposição à fumaça ambiental do tabaco (FAT) e infecção pelo H. pylori sobre a incidência de amigdalite crônica em crianças chinesas. TIPO DE ESTUDO E LOCAL: Estudo transversal desenvolvido num ambulatório na China. MÉTODOS: Pacientes pediátricos com amigdalite crônica foram recrutados. A infecção por H. pylori foi determinada segundo a presença de anticorpos H. pylori CagA IgG. Foi determinado o nível de cotinina sérica e exposição à FAT de todos os participantes. RESULTADOS: Não houve diferença significativa entre crianças com amigdalite crônica na infecção por H. pylori e sem amidalite, mas existia diferença significativa na FAT entre os dois grupos (P = 0,011). Em seguida, estudamos a associação entre FAT e amigdalite crônica com base no status de infecção por H. pylori. Nos pacientes com infecção por H. pylori, houve diferença significativa na distribuição de FAT entre os grupos de amigdalite crônica e controle (P = 0,022). Tomando os participantes sem FAT como referência, a análise de regressão logística multivariada mostrou que aqueles com alta FAT tinha maior susceptibilidade à amigdalite crônica (OR ajustado IC = 2,33, 95%: 1,67-3,25, ajustado P < 0,001). No entanto, naqueles sem infecção por H. pylori, a FAT não predispôs a amigdalite crônica. CONCLUSÃO: Nossos achados sugerem que a exposição ao tabaco é um fator de risco para amigdalite crônica em crianças com infecção por H. pylori.

Lack of association between helicobacter pylori's virulence and increased serum c-reactive protein levels in functional dyspeptic patients

ABSTRACT Background Recently, a great variety of studies aimed to investigate and even suggestHelicobacter pylori as an important key factor in gastrointestinal and non-gastrointestinal events development. The well-established relationship between bacterial virulence and increased risk for peptic ulcer or gastric carcinoma is not so clear when comparing inflammation markers alterations, such C-reactive protein, with the pathogen. Objective The objective of this study was to evaluate the presence of H. pylori, bacterial virulence and C-reactive protein serum levels in individuals diagnosed with functional dyspepsia. Methods Were prospectively included in this study 489 dyspeptic individuals. They fulfill Rome III clinical criteria for the diagnosis of functional dyspepsia with no organic disease at endoscopy. The bacterial infection was established by histology and urease rapid test. The levels of serum C-reactive protein were obtained by immunonefelometry and CagA status ofH. pylori positive individuals was determined through an imunoenzimatic assay. Results Prevalence rate of H. pylori was 66.3% and virulence factor CagA was detected in nearly 43% of positive samples. In addition, it has been noticed an association between Ilex paraguariensis(yerba maté) consumption and pathogen's prevalence. An important effect of bacterial infection on inflammation was only observed in gastric epithelium. Conclusion No systemic response to the pathogen, measured through C-reactive protein levels, was observed, regardless of CagA status. Otherwise, the intake of yerba maté should be considered as a cultural factor possibly related toH. pylori's transmission.

RESUMO Contexto Recentemente, uma grande variedade de estudos tem investigado e até mesmo sugerido a presença de Helicobacter pylori como um importante fator no desenvolvimento de eventos restritos ou não ao trato gastrointestinal. A relação já bem estabelecida entre virulência bacteriana e risco aumentado para úlcera péptica ou adenocarcinoma gástrico não parece estar tão elucidada quando se comparam alterações de marcadores inflamatórios, como a proteína C-reativa, com a presença do patógeno. Objetivo O objetivo deste estudo foi avaliar a presença da infecção por H. pylori, a virulência bacteriana e os níveis séricos de proteína C-reativa em indivíduos diagnosticados com dispepsia funcional. Métodos Foram incluídos neste estudo, prospectivamente, 489 indivíduos dispépticos. Os pacientes deveriam preencher os critérios clínicos de Roma III para o diagnóstico de dispepsia funcional sem apresentar doença orgânica evidenciada a partir da endoscopia. A infecção bacteriana foi estabelecida por histologia e pelo teste rápido da urease. Os níveis de proteína C-reativa foram quantificados através de imunonefelometria e o status para a presença da CagA dos indivíduos infectados por H. pylorifoi determinado por ensaio imunoenzimático. Resultados A taxa de prevalência de H. pylori foi de 66.3% e o fator de virulência CagA foi detectado em aproximandamente 43% das amostras positivas. Adicionalmente, denotou-se uma associação entre o consumo deIlex paraguariensis (chimarrão) e a prevalência do patógeno. Um importante efeito da infecção bacteriana na inflamação apenas foi observado localmente, no epitélio gástrico. Conclusão Não foi evidenciada resposta sistêmica ao patógeno aferido através dos níveis de proteína C-reativa, independentemente do status para CagA. Por outro lado, o consumo de chimarrão pode ser sugerido como um fator cultural possivelmente relacionado à transmissão de H. pylori.

Mycobacterium tuberculosis epitope-specific interferon-g production in healthy Brazilians reactive and non-reactive to tuberculin skin test

The interferon (IFN)-γ response to peptides can be a useful diagnostic marker of Mycobacterium tuberculosis (MTB) latent infection. We identified promiscuous and potentially protective CD4+ T-cell epitopes from the most conserved regions of MTB antigenic proteins by scanning the MTB antigenic proteins GroEL2, phosphate-binding protein 1 precursor and 19 kDa antigen with the TEPITOPE algorithm. Seven peptide sequences predicted to bind to multiple human leukocyte antigen (HLA)-DR molecules were synthesised and tested with IFN-γ enzyme-linked immunospot (ELISPOT) assays using peripheral blood mononuclear cells (PBMCs) from 16 Mantoux tuberculin skin test (TST)-positive and 16 TST-negative healthy donors. Eighty-eight percent of TST-positive donors responded to at least one of the peptides, compared to 25% of TST-negative donors. Each individual peptide induced IFN-γ production by PBMCs from at least 31% of the TST-positive donors. The magnitude of the response against all peptides was 182 ± 230 x 106 IFN-γ spot forming cells (SFC) among TST-positive donors and 36 ± 62 x 106 SFC among TST-negative donors (p = 0.007). The response to GroEL2 (463-477) was only observed in the TST-positive group. This combination of novel MTB CD4 T-cell epitopes should be tested in a larger cohort of individuals with latent tuberculosis (TB) to evaluate its potential to diagnose latent TB and it may be included in ELISPOT-based IFN-γ assays to identify individuals with this condition.

The first report of the vanC1 gene in Enterococcus faecium isolated from a human clinical specimen

The vanC1 gene, which is chromosomally located, confers resistance to vancomycin and serves as a species marker for Enterococcus gallinarum. Enterococcus faecium TJ4031 was isolated from a blood culture and harbours the vanC1gene. Polymerase chain reaction (PCR) assays were performed to detect vanXYc and vanTc genes. Only the vanXYc gene was found in the E. faecium TJ4031 isolate. The minimum inhibitory concentrations of vancomycin and teicoplanin were 2 µg/mL and 1 µg/mL, respectively. Real-time reverse transcription-PCR results revealed that the vanC1and vanXYc genes were not expressed. Pulsed-field gel electrophoresis and southern hybridisation results showed that the vanC1 gene was encoded in the chromosome. E. faecalis isolated from animals has been reported to harbour vanC1gene. However, this study is the first to report the presence of the vanC1gene in E. faecium of human origin. Additionally, our research showed the vanC1gene cannot serve as a species-specific gene of E. gallinarum and that it is able to be transferred between bacteria. Although the resistance marker is not expressed in the strain, our results showed that E. faecium could acquire the vanC1gene from different species.

Cloning and expression of a recombinant CagA -gene fragment of Helicobacter pylori and its preliminary evaluation in serodiagnosis / Clonación y expresión de un fragmento recombinante del gen cagA de Helicobacter pylori y su evaluación preliminar en el serodiagnóstico

Introduction: Helicobacter pylori strains expressing cytotoxic CagA protein are more commonly associated with peptic ulceration, atrophic gastritis and gastric adenocarcinoma than those lacking CagA. Determination of anti-CagA antibodies, therefore, acquires a relevant clinical significance in the serological detection of H. pylori infection and disease risk prediction. However, the CagA-serology has been questioned due to the differences found in their performance evaluations in different populations. Objective: To obtain a recombinant CagA fragment useful for serodiagnosis of H. pylori infection Methods: A fragment of the cagA gene was cloned into a prokaryotic T7 RNA polymerase expression vector. A recombinant C-terminal His 6 -tagged CagA was expressed, subsequently solubilized with urea and purified by immobilized metal affinity chromatography. The performance of the recombinant protein was evaluated using 180 human serum samples with an in-house Western blot assay compared to the Helicoblot 2.1 reference test. Results: The expressed His 6 -tagged CagA showed an immunoreactive 80kDa band as was revealed by SDS-PAGE and Western blot analysis using two different specific anti-CagA polyclonal antibodies. The recombinant protein was successfully purified obtaining a 93% of purity. The performance analysis of the purified recombinant antigen showed good immunoreactivity and exhibited values of sensitivity, specificity and accuracy of 88.1%, 100% and 92.7%, respectively. Conclusion: The CagA fragment of the study may constitute a useful tool for serological diagnosis of CagA-positive H. pylori infection.

Introducción. Las cepas de Helicobacter pylori que expresan la citotoxina CagA, se asocian más frecuentemente con úlcera péptica, gastritis atrófica y adenocarcinoma gástrico que las que carecen de esta citotoxina. Por lo anterior, el determinar la presencia de anticuerpos anti-CagA adquiere gran importancia clínica en la detección serológica de la infección por H. pylori y la predicción del riesgo de enfermedades. Sin embargo, los métodos serológicos que emplean CagA han sido cuestionados debido a las diferencias encontradas en las evaluaciones de su desempeño en diversas poblaciones. Objetivo. Obtener un fragmento recombinante de la proteína CagA para el serodiagnóstico de la infección por H. pylori . Materiales y métodos. Un fragmento del gen cagA fue clonado en un vector de expresión procariota que contenía el promotor de la T7 ARN polimerasa. El fragmento de la proteína CagA con seis histidinas en la región C-terminal, se expresó, se solubilizó con urea y se purificó por cromatografía de afinidad con iones metálicos inmovilizados. El desempeño de la proteína recombinante se evaluó empleando un método in house de Western Blot y 180 sueros humanos. Los resultados se compararon con la prueba de referencia Helicoblot 2.1. Resultados. La proteína CagA expresada mostró una banda inmunorreactiva de 80 kDa en el Western Blot al emplear dos anticuerpos policlonales anti-CagA específicos. La proteína recombinante fue purificada hasta un 93 % de pureza y el análisis de desempeño del antígeno recombinante purificado mostró buena inmunorreacción y exhibió valores de sensibilidad, especificidad y exactitud de 88,1 %, 100 % y 92,7 %, respectivamente. Conclusiones. El fragmento de la proteína CagA del estudio puede constituir una herramienta útil para el diagnóstico serológico de la infección por cepas de H. pylori positivas para CagA.

Identification of serological biomarkers of infection, disease progression and treatment efficacy for leprosy

Although leprosy is curable with drug treatment, the identification of biomarkers of infection, disease progression and treatment efficacy would greatly help to reduce the overall prevalence of the disease. Reliable biomarkers would also reduce the incidence of grade-2 disability by ensuring that those who are most at risk are diagnosed and treated early or offered repeated treatments in the case of relapse. In this study, we examined the reactivity of sera from lepromatous and tuberculoid leprosy patients (LPs) against a panel of 12 recombinant Mycobacterium leprae proteins and found that six proteins were strongly recognised by multibacillary (MB) patients, while only three were consistently recognised by paucibacillary patients. To better understand the dynamics of patient antibody responses during and after drug therapy, we measured antibody titres to four recombinant proteins, phenolic glycolipid-I and lipoarabinomannan at baseline and up to two years after diagnosis to investigate the temporal changes in the antibody titres. Reactivity patterns to individual antigens and decreases in antibody titres were patient-specific. Antibody titres to proteins declined more rapidly vs. those to carbohydrate and glycolipid antigens. Compared to baseline values, increases in antibody titres were observed during reactional episodes in one individual. Additionally, antibody responses against a subset of antigens that provided a good prognostic indicator of disease progression were analysed in 51 household contacts of MB index cases for up to two years. Although the majority of these contacts showed no change or exhibited decreases in antibody titres, seven individuals developed higher titres towards one or more of these antigens and one individual with progressively higher titres was diagnosed with borderline lepromatous leprosy 19 months after enrolment. The results of this study indicate that antibody titres to specific M. leprae antigens can be used to monitor treatment efficacy in LPs and assess disease progression in those most at risk for developing this disease.

Serologic follow-up of IgG responses against recombinant mycobacterial proteins ML0405, ML2331 and LID-1 in a leprosy hyperendemic area in Venezuela

Leprosy is a slowly evolving disease that occurs mainly in adults. In this study, the Mamaría Village, state of Portuguesa was selected because it had one of the highest prevalence rates (13.25%) of leprosy cases in 1997. Between 1998-2004, 20.2% of the 89 cases registered in this village were less than 15 years old and 61.8% were males. Pau-cibacillary (PB) lesions were the predominant clinical forms identified, although also multibacillary (MB) forms were found. Additionally, 76% of the patients were bacteriologically negative. At the time of diagnosis, 75% of the patients presented with grade 0 disabilities, 23% with grade 1 and 2% with grade 2. Serum samples were collected from 18 PB and 15 MB patients, in addition to 14 family contacts, at the beginning and end of treatment. All the groups were re-evaluated during a three-year period (2008-2011). The proteins used for evaluation were ML0405, ML2331 and LID-1. These mycobacterial proteins were highly specific for Mycobacterium leprae and the IgG responses decreased in both MB and PB patients during multidrug treatment. Our results suggest that these antigens could be used as markers for successful treatment of non-reactional lepromatous patients.

Seroreactivity to new Mycobacterium leprae protein antigens in different leprosy-endemic regions in Brazil

New Mycobacterium leprae protein antigens can contribute to improved serologic tests for leprosy diagnosis/classification and multidrug therapy (MDT) monitoring. This study describes seroreactivity to M. leprae proteins among participants from three highly endemic leprosy areas in Brazil: central-western Goiânia/Goiás (GO) (n = 225), Rondonópolis/Mato Grosso (MT) (n = 764) and northern Prata Village/Pará (PA) (n = 93). ELISA was performed to detect IgG to proteins (92f, 46f, leprosy IDRI diagnostic-1, ML0405, ML1213) and IgM to phenolic glycolipid-I (PGL-I). Multibacillary (MB) leprosy had positive rates for PGL-I that were similar to those for proteins; however, some anti-PGL-I-negative subjects were positive for proteins, suggesting that adding protein antigen to PGL-I can enhance the sensitivity of MB leprosy detection. In MT, different degrees of seroreactivity were observed and ranked for MB, former patients after MDT, paucibacillary (PB) leprosy, household contact (HHC) and endemic control (EC) groups. The seroreactivity of PB patients was low in GO and MT. HHCs from different endemic sites had similar IgG antibody responses to proteins. 46f and 92f were not recognised by most tuberculosis patients, ECs or HHCs within GO, an area with high BCG vaccination coverage. Low positivity in EC and HHC was observed in PA and MT. Our results provide evidence for the development of an improved serologic test that could be widely applicable for MB leprosy testing in Brazil.

Identificación bacteriana basada en el espectro de masas de proteínas: Una nueva mirada a la microbiología del siglo XXI / Bacterial identification based on protein mass spectrometry: A new insight at the microbiology of the 21st century

Bacterial identification is important for the proper treatment of infected patients hospitalized with serious infections especially in critical care units. Identification by conventional methods used in microbiology laboratories takes at least 16 hours since a culture is positive. The introduction of mass spectrometry, specifically MALDI-TOF MS (matrix-assisted laser desorption/ ionization time-of-flight mass spectrometer) in the microbiology laboratory could mean a radical change in the identification accuracy, turn around time (6 minutes per bacteria) and cost (about 5 times cheaper than conventional identification). Since its introduction in clinical microbiology laboratories in 2008, many reports about its usefulness in identifying microorganisms from colonies, as well as directly from positive blood cultures and urine samples have been published. This review describes MALDI-TOF MS methodology, its identification performance for bacteria (aerobic and anaerobic), mycobacterium and yeasts, its future applications in microbiology and its main disadvantages.

La identificación bacteriana es muy importante en el manejo adecuado de los pacientes infectados, especialmente aquellos con infecciones graves hospitalizados en unidades de pacientes críticos. La identificación por los métodos convencionales utilizados en los laboratorios de microbiología clínica demora al menos 16 horas desde que un cultivo es positivo. La introducción de la espectrometría de masas, específicamente del espectrómetro de masas por tiempo de migración (tiempo de vuelo) con desorción/ionización laser asistida por una matriz (MALDI-TOF MS, por su sigla en inglés matrix-assisted laser desorption/ionization time-of-flight mass spectrometer), en el laboratorio de microbiología podría significar un cambio radical en la precisión de la identificación, el tiempo de detección (6 minutos por bacterias) y el costo (aproximadamente 5 veces más económico que la identificación convencional). Desde su introducción en los laboratorios de microbiología clínica en el año 2008, se han escrito numerosas publicaciones sobre su utilidad en la identificación de microorganismos desde colonias, así como directamente desde hemocultivos positivos y de muestras de orina. Esta revisión describe la metodología de MALDI-TOF MS, su rendimiento en la identificación de bacterias aerobias, anaerobias, micobacterias y levaduras, sus futuras aplicaciones en microbiología y sus principales desventajas.

Seroreactivity to new Mycobacterium leprae protein antigens in different leprosy-endemic regions in Brazil

New Mycobacterium leprae protein antigens can contribute to improved serologic tests for leprosy diagnosis/classification and multidrug therapy (MDT) monitoring. This study describes seroreactivity to M. leprae proteins among participants from three highly endemic leprosy areas in Brazil: central-western Goiânia/Goiás (GO) (n = 225), Rondonópolis/Mato Grosso (MT) (n = 764) and northern Prata Village/Pará (PA) (n = 93). ELISA was performed to detect IgG to proteins (92f, 46f, leprosy IDRI diagnostic-1, ML0405, ML1213) and IgM to phenolic glycolipid-I (PGL-I). Multibacillary (MB) leprosy had positive rates for PGL-I that were similar to those for proteins; however, some anti-PGL-I-negative subjects were positive for proteins, suggesting that adding protein antigen to PGL-I can enhance the sensitivity of MB leprosy detection. In MT, different degrees of seroreactivity were observed and ranked for MB, former patients after MDT, paucibacillary (PB) leprosy, household contact (HHC) and endemic control (EC) groups. The seroreactivity of PB patients was low in GO and MT. HHCs from different endemic sites had similar IgG antibody responses to proteins. 46f and 92f were not recognised by most tuberculosis patients, ECs or HHCs within GO, an area with high BCG vaccination coverage. Low positivity in EC and HHC was observed in PA and MT. Our results provide evidence for the development of an improved serologic test that could be widely applicable for MB leprosy testing in Brazil

On improving assessment of in-hospital mortality and ICU admission in community-acquired pneumonia patients using the eCURB

Assessment of severity of the disease in community-acquired pneumonia [CAP] is very important to decide the site of care. The conventional CURB-65 score is composed of five separate elements namely, Confusion, Uremia, Respiratory rate, BP, and age >/=65 years. These elements could be calculated electronically. The electronic CURB [eCURB] utilizes the 5 CURB-65 data elements as continuous, weighted variables. The aim of this study was to evaluate the performance of eCURB elements in predicting in-hospital mortality and ICU admission in comparison to the conventional CURB-65. This study was conducted upon 134 adult patients diagnosed as CAP and confirmed by radiographic findings, admitted to chest department, Assiut University Hospital, Egypt. The CURB-65 elements were retrospectively extracted from the medical records. The eCURB variables were introduced to electronically calculate the risk using the Excel appendix model [provided by Prof. Nanthan Dean, University of Utah, Salt Lake city, USA] and its predictive values and area under the receiver-operating characteristic [ROC] curve were compared with the conventional CURB-65 in predicting in-hospital mortality and the need for ICU admission. The study revealed that the conventional CURB-65 score could predict in-hospital mortality with an area under the curve [AUC] of 0.81 and the need for ICU admission with an AUC of 0.87. Using the eCURB-65 elements proved to be superior to the conventional CURB-65 in predicting in-hospital mortality with cut off point > 7.5 and an AUC of 0.83 [P < 0.0001]. Also, eCURB was better than conventional CURB-65 in predicting ICU admission with cut off point > 3.8 and an AUC of 0.89 [P < 0.0001]. Using the eCURB proved to be a valuable tool in predicting in-hospital mortality and ICU admission in patients with CAP with a significant superiority over conventional CURB-65 in both variables. Further prospective studies on a larger cohort are recommended

Detection of fluoroquinolone resistance in mycobacterium tuberculosis clinical isolates as determined by gyrA / B gene mutation by using PCR technique

Fluoroquinolones are broad-spectrum antimicrobial agents that have been used with increasing frequency over the past decade. Fluoroquinolones have in vitro and in vivo activity against Mycobacterium tuberculosis. However, resistance to fluoroquinolones in cases of tuberculosis is not routinely assessed. Mutations in a small region of gyrA, called quinolone resistance-determining region [QRDR] and, less frequently, in gyrB are the primary mechanism of FQ resistance in M. tuberculosis. PCR-based techniques provide new possibilities for the rapid diagnosis of first- and second-line drug resistance. There were 40 consecutive adults, who had culture confirmed pulmonary tuberculosis during the study period. Mutations were observed in the QRDRs of both gyrA and gyrB in 22 isolates [55%]. Only gyrA +ve in 7[17.5%] isolates. Only gyrB +ve in 5[12.5%] isolates. Total gyrA +ve in 29[72.5%] and total gyrB +ve in 28[70%] isolates. Both gyrA and gyrB -ve in 6 [15%]. The incidence of FO-resistant M. tuberculosis is gradually increasing to alarming levels this may be due to wide spread use of this vital groups of drugs in community-acquired pneumonia and urinary tract infections

Prognosis of Korean patients with primary biliary cirrhosis / 대한간학회지

No abstract available.

Clinical features and prognosis of primary biliary cirrhosis in Korea / 대한간학회지

BACKGROUND/AIMS: This study investigated the clinical features and prognosis of primary biliary cirrhosis (PBC) in Korea. METHODS: Clinical data of patients diagnosed as PBC between 1997 and 2008 at eight referral hospitals were analyzed retrospectively. PBC was diagnosed based on liver function tests, presence of serum antimitochondrial antibody (AMA), and histopathological findings. RESULTS: In total, 251 patients (218 females, 33 males; mean age 54 years) were enrolled, and the mean follow-up duration was 33.5 months. At the diagnosis, 61% of the patients were asymptomatic, 12% had decompensated liver cirrhosis, and 98% were positive for AMA. The serum alkaline phosphate (ALP) level was 2.6 times the upper limit of normal, aspartate aminotransferase was 105 U/L, and bilirubin was 2.0 mg/dL. The mean Mayo risk score was 5.5, and the Child-Pugh class was A, B, and C in 79%, 19%, and 2% of the patients, respectively. Ursodeoxycholic acid (UDCA) was used for treatment in 88% of the patients, among which 70% exhibited biochemical responses defined as normalization or a >40% decrease in ALP at 6 months. Eight deaths occurred during the follow-up; the causes were variceal bleeding, hepatic failure, and sepsis. The overall 5-year survival rate was 95%. The poor prognostic factors were being older than 60 years, high bilirubin, low albumin, ascites, high Mayo risk score, Child-Pugh class C, and initial presence of hepatic decompensation. CONCLUSIONS: Most patients diagnosed as PBC were asymptomatic, and these patients had a favorable short-term prognosis. The prognosis of PBC was dependent on the initial severity of liver disease.

Helicobacter pylori em crianças e associação de cepas CagA na transmissão mãe-filho na Amazônia brasileira / Helicobacter pylori in children and association with CagA strains in mother-child transmission in the Brazilian Amazon region

Investigou-se a prevalência de infecção pela Helicobacter pylori em amostras de sangue de 100 crianças de 1 a 12 anos e de suas mães através dos métodos de hemaglutinação indireta e anti-CagA pelo ensaio ELISA. Destas 100 crianças, foram obtidas 79 amostras de fezes e realizada pesquisa de antígenos da bactéria nas fezes por ELISA de captura. Os antígenos foram detectados em 54,4 por cento (43/79) das crianças, e os anticorpos no soro em 43 por cento (34/79), métodos que apresentaram desempenhos semelhantes, com maiores discordâncias nas crianças de 1 a 4 anos. A soroprevalência nas crianças foi de 50 por cento (50/100) e nas mães de 86 por cento (86/100). Mães infectadas representaram fator de risco 19 vezes superior ao de mães soronegativas para determinar infecção em seus filhos (p < 0,05), sobretudo as mães com cepas CagA+ (p < 0,05). O contato direto pessoa-pessoa pode ser um modo de transmissão desta infecção.

The prevalence of Helicobacter pylori infection was investigated in blood samples from 100 children aged 1 to 12 years and from their mothers, by means of the indirect hemagglutination and anti-CagA methods, using ELISA assays. From these 100 children, 79 stool samples were obtained and bacterial antigens in the stools were investigated using capture ELISA. The antigens were detected in 54.4 percent (43/79) of the children, and serum antibodies in 43 percent (34/79). These methods presented similar performance, with greatest disagreement among the children aged 1 to 4 years. The seroprevalence was 50 percent (50/100) among the children and 86 percent (86/100) among the mothers. Infected mothers represented a risk factor that was 19 times greater than that of seronegative mothers, with regard to infecting their children (p < 0.05), especially the mothers with CagA+ strains (p < 0.05). Direct person-to-person contact may be a transmission method for this infection.

Can anti-Helicobacter pylori and anti-CagA antibodies be used to select patients with dyspepsia for gastroscopy?

BACKGROUND: agA IgG antibody in sera may indicate presence of peptic ulcer disease among dyspeptic patients and therefore may be used as a serological marker to identify high risk patients for peptic ulcer who can be subjected to endoscopy. Present study was performed to identify association of CagA IgG antibody in patients with peptic ulcer. METHODS: Consecutive patients with dyspepsia were subjected to endoscopy and sera was collected from each. Rapid urease test in antral tissue collected from each patient by endoscopic biopsy was performed. Antral tissue was also examined histologically. IgG Antibody against H. Pylori and CagA IgG antibody was tested in each patients sera. RESULTS: Out of 82 patients with dyspepsia included in the study 28 had peptic ulcer. Of whom 26 were positive for anti IgG H. Pylori antibody. More than 80% patients with peptic ulcer patients had detectable anti Cag A antibody in contrast to 33% patients with non ulcer dyspepsia (P < 0.001). CONCLUSION: Anti-Cag A antibody may be used as a screening test in patients with dyspepsia to select high risk patients for peptic ulcer for upper gastrointestinal endoscopy.

Comparison of two methods to detect carbapenemase & metallo-beta- lactamase production in clinical isolates.

BACKGROUND & OBJECTIVE: Bacterial resistance has greatly hampered effective treatment of patients in clinical settings. Non-fermenting Gram-negative bacilli (NFGNB) are common nosocomial pathogens. In this study we attempted to develop a convenient test for early detection of carbapenemase and metallo-beta-lactamase (MbetaL) production in NFGNB. Lack of sufficient reports from India in this area indicated the need for this study. METHODS: A total of 50 imipenem resistant NFGNB were speciated, and their resistance reconfirmed by disk diffusion and minimum inhibitory concentration (MIC) determination by agar dilution. Two different methods namely modified Hodge and EDTA disk synergy tests were evaluated for carbapenemase and metallo-beta-lactamase (MbetaL) production. RESULTS: Of the 50 imipenem resistant NFGNB, 48 and two respectively fell in the resistant and intermediate range in MIC using agar dilution. Majority of these were Pseudomonas aeruginosa (n=28), followed by Burkholderia cepacia (n=9). The modified Hodge test could detect 28 strains as carbapenemase and MbetaL producers, while the EDTA disk synergy test was able to detect an additional 8 strains producing MbetaL and carbapenemase. INTERPRETATION & CONCLUSION: Pseudomonas aeruginosa was found to be the predominant NFGNB in our hospital setting and EDTA disk synergy could detect more carbapenemase and metallo-beta- lactamase producers compared to modified Hodge test.

Prevalence of CagA and VacA genotype of Helicobacter pylori in Thai children.

Seroprevalence of Helicobacter pylori infection in children is variable according to geographical location and family sanitation. A previous study in Bangkok showed an incidence of 25.5% in 1998. The higher incidence in the urban and rural area is predicted in lower economic classes and poor sanitation. OBJECTIVE: To study the occurance of CagA and VacA genotype in Thai children using the Western blot technique. MATERIAL AND METHOD: Sera of 159 Thai native children aged 0-15 year without associated abdominal pain from different provinces in 4 parts of the Kingdom of Thailand were tested with the rapid screening test for H. pylori. The positive specimen was further tested with the Western blot technique for determination of Urea A (p37), CagA (p116) and VacA (p89). RESULT: Fiftyfive andfifty two (34.6%) were tested positive by the rapid test while 32.7% were positivefor the band of current infection marker (CIM). The 28 selected positive sera with complete history of housing and water supply were analysed. Thai children living in urban areas have a higher prevalence and the CagA+, VacA+ are found in 96.43% of infected patients. The transmission may be through the water supply. CONCLUSION: A high prevalence of Helicobacter pylori infection was found in childhood period in urban areas and may be associated with the local water supply.

Seroprevalencia de anticuerpos anti-caga de helicobacter pylori y su relacion con factores epidemiologicos wn Santa Fe / Seroprevalence of helicobacter pylori anti-caga antibodies and its relationship with epidemiologic factors in Santa Fe

La detección precoz de anticuerpos anti-CagA en adultos jóvenes tendría gran impacto clínico en la prevención del cáncer gástrico. El objetivo del presente trabajo fue determinar en nuestra región la seroprevalencia de los anticuerpos IgG y anti-CagA de Helicobacter pylori, con una técnica no invasiva y fácil de realizar; valorando su relación con diferentes factores de riesgos epidemiológicos. Se incorporaron 435 voluntarios de diferentes centros de salud con edad promedio de 40 años. Mediante una encuesta personalizada se registraron variables demográficas, condiciones socieconómicas, entre otros datos de interés. En el suero de los individuos se determinó la presencia de anticuerpos IgG específicos y anti-CagA del Helicobacter pylori mediante una técnica de enzimoinmunoensayo. La prevalencia de los anticuerpos IgG fue de 52.2%, siendo positivo en 152 (53.7%) mujeres y 75 (49%) varones. Los anticuerpos IgG estaban presentes en el 65.0% de los individuos con síntomas y en el 44.1% de los asintomáticos. La prevalencia de los anticuerpos anti-CagA en la muestra estudiada fue 33.1%, presentándose en el 63.4% de los sujetos seropositivos (IgG), siendo positivo en 96 (33.9%) mujeres y 48 (31.6%) varones. La prevalencia fue 45.4% y 25.7% en los sintomáticos y asintomáticos respectivamente. Se demostró que los anticuerpos IgG están asociados a la edad, zona de residencia, nivel educacional y número de dormitorios por vivienda; mientras que los anticuerpos anti-CagA dependen de la zona de residencia y de la presencia de síntomas. La influencia de la variable predictiva síntomas sobre la presencia de los anticuerpos anti-CagA revelan la importancia selectiva en la corroboración de parámetros clínicos de las enfermedades gastroduodenales asociadas a Helicobacter pylori.