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1.
Rev. Soc. Bras. Clín. Méd ; 18(4): 196-199, DEZ 2020.
Artigo em Português | LILACS | ID: biblio-1361597

RESUMO

Objetivo: Analisar a acurácia do teste rápido da urease para detecção de Helicobacter pylori comparado com o exame histopatológico. Métodos: Estudo prospectivo e descritivo realizado de abril de 2018 a maio de 2019 em um Serviço de Endoscopia e Biliopancreática e em um laboratório de patologia. A amostra foi composta de 64 pacientes, de ambos os sexos, com idade de 35 a 81 anos, que apresentavam queixas dispépticas. Foram realizados exame histopatológico e teste rápido da urease. Os dados foram analisados pelo R Core team 2019 e submetidos a análises descritivas (variáveis categóricas) e inferenciais (teste de associação de qui-quadrado de Pearson e teste de Mann-Whitney). O nível de significância adotado foi de 5%. Resultados: O teste rápido da urease demonstrou que dez pacientes foram verdadeiros-positivos, 39 verdadeiros-negativos, três falsos-positivos, 12 falsos-negativos, com sensibilidade de 45,4% (25,1% a 67,3%), especificidade de 92,9% (79,4% a 98,1%), valor preditivo positivo de 76,9% (45,9% a 93,8%), valor preditivo negativo de 76,5% (62,2% a 86,7%), acurácia de 76,6% (64,0% a 85,9%), razão de chance diagnóstica 10,8 (2,56 a 45,9), índice de Youden 0,38 (0,16 a 0,60) e taxa de erro de 23,4% (14,1% a 36,0%). Conclusão: O teste rápido da urease apresentou baixa capacidade de detectar pacientes infectados, menor acurácia em relação ao estudo anatomopatológico e alta especificidade. O teste pode ser útil no momento da realização da endoscopia, por fornecer resultado rápido e barato para detectar H. pylori. O diagnóstico da bactéria apresenta maior confiabilidade com a realização dos dois métodos para pesquisa de H. pylori.


Objective: To analyze the accuracy of the rapid urease test for Helicobacter pylori detection when compared with the histopathological examination. Methods: This is a prospective and descriptive study conducted from April 2018 to May 2019, at an Endoscopy and Biliopancreatic Service and in a pathology laboratory. The sample consisted of 64 male and female patients aged 35 to 81 years old with dyspeptic complaints. Histopathological examination and rapid urease test were performed. Data were analyzed by R Corel team 2019 and underwent descriptive (categorical variables) and inferential (Pearson's Chi-squared association test and Mann-Whitney test) analyzes. The significance level adopted was 5%. Results: The rapid urease test showed that ten patients were true positive, 39 true negative, three false-positive, and 12 false-negative, and sensitivity was of 45.4% (25.1% to 67.3%), specificity 92.9% (79.4% to 98.1), positive predictive value of 76.9% (45.9-93.8%), negative predictive value of 76.5% (62.2% to 86.7%), accuracy of 76.6% (64.0% to 85.9%), diagnostic odds ratio of 10.8 (2.56% to 45.9), Youden index 0.38 (0.38% to 0.60), and error rate 23.4 (14.1% to 36.0%). Conclusion: The rapid urease test showed low ability to detect infected patients, lower accuracy compared to the pathological study, and high specificity. The test may be useful at the time of endoscopy, as it provides a quick and inexpensive result to detect H. pylori. The diagnosis of the bacterium is more reliable when both methods for H. pylori investigation are performed


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Urease/análise , Helicobacter pylori/enzimologia , Infecções por Helicobacter/diagnóstico , Mucosa Gástrica/patologia , Biópsia , Estudos Prospectivos , Sensibilidade e Especificidade , Gastroscopia , Dispepsia/diagnóstico
2.
Rev. argent. microbiol ; 50(4): 359-364, Dec. 2018. ilus, tab
Artigo em Inglês | LILACS | ID: biblio-977257

RESUMO

Helicobacter pylori is a gastric pathogen that is widely recognized as a causative agent of gastric disease. Its eradication is variable, mainly due to increased resistance to clarithromycin. Our objective was: to evaluate (i) if the biopsy specimen used for the rapid urease test is a useful sample to detect resistance to clarithromycin by PCR-RFLP and (ii) the distribution of A2142G and A2143G point mutations in the 23S rRNA gene, in relation to virulence factors in our region. Gastric specimens were collected from adult dyspeptic patients (n = 141) and H. pylori was investigated by the rapid urease test, histopathological analysis and PCR for the hsp60 gene. Clarithromycin resistance was detected by PCR-RFLP in 62 H. pylori (+) paired biopsy specimens submitted to molecular analysis and the rapid urease test. H. pylori virulence factors were analyzed by multiplex PCR using specific primers for the cagA, vacA and babA2 genes. Thirteen out of 62 strains (20.9%) were resistant to clarithromycin: 6/13 (46.2%) harbored the A2143G mutation whereas 7/13 (53.8%) carried the A2142G point mutation. vacA m1s1 was the most frequent genotype among the resistant strains. In conclusion, the biopsy specimens used for the rapid urease test were suitable samples for clarithromycin resistance detection in patients infected with H. pylori, which became especially useful in cases where the number or size of the biopsies is limited. In addition, this is the first report of a molecular analysis for clarithromycin resistance performed directly from gastric biopsies in our region.


Helicobacter pylori es un patógeno ampliamente reconocido como causante de enfermedad gástrica. Su erradicación es variable, principalmente debido al incremento de la resistencia a claritromicina. Nuestros objetivos fueron evaluar la utilidad de la biopsia usada para realizar el test rápido de ureasa en la detección de resistencia a claritromicina por PCR-RFLP y conocer la distribución de las mutaciones puntuales A2142G y A2143G en el gen ARNr 23S, en relación con los factores de virulencia en nuestra región. Se recolectaron muestras gástricas (n=141) provenientes de pacientes adultos dispépticos y se investigó la presencia de H. pylori mediante el test rápido de ureasa, análisis histopatológico y PCR para el gen hsp60. La resistencia a claritromicina se analizó por PCR-RFLP en 62 muestras pareadas de biopsias gástricas H. pylori+ destinadas al análisis molecular y al test rápido de ureasa. Los factores de virulencia de H. pylori fueron analizados mediante PCR multiplex usando oligonucleótidos específicos para los genes cagA, vacA y babA2. Trece de 62 cepas (20,9%) fueron resistentes a claritromicina, 6/13 (46,2%) llevaron la mutación A2143G, mientras que 7/13 (53,8%) presentaron la mutación A2142G. El genotipo vacA s1m1 fue el más frecuente entre las cepas resistentes a claritromicina. En conclusión, las biopsias destinadas al test rápido de ureasa fueron muestras apropiadas para la detección de la resistencia a claritromicina en pacientes infectados con H. pylori. Esto es especialmente útil en aquellos casos en los que el número o el tamaño de las muestras son limitados. Además, este es el primer reporte de estudio de resistencia a claritromicina (mediante técnicas moleculares), directamente de biopsias gástricas en nuestra región.


Assuntos
Humanos , Helicobacter pylori/efeitos dos fármacos , Infecções por Helicobacter/diagnóstico , Claritromicina/farmacologia , Fatores de Tempo , Urease/metabolismo , Polimorfismo de Fragmento de Restrição , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Helicobacter pylori/enzimologia , Helicobacter pylori/genética , Helicobacter pylori/patogenicidade , Infecções por Helicobacter/microbiologia , Mutação Puntual , Farmacorresistência Bacteriana , Testes Diagnósticos de Rotina/métodos
3.
Rev. gastroenterol. Perú ; 37(1): 53-57, ene.-mar. 2017. ilus, tab
Artigo em Espanhol | LILACS | ID: biblio-991224

RESUMO

Objetivos: Validar un test rápido de la ureasa (TRU) en el Hospital Cayetano Heredia (HCH) de Lima, Perú Materiales y métodos: Estudio observacional prospectivo. Se incluyó 181 pacientes mayores de 18 años de edad con síntomas dispépticos, que fueron sometidos a endoscopía digestiva alta en el Servicio de Gastroenterología del HCH y que no hubiesen recibido durante las últimas cuatro semanas inhibidores de la bomba de protones (IBPs), bismuto o antibióticos. Se tomó dos biopsias de antro una para hacer el TRU (Sensibacter pylori test®) y otra para anatomía patológica con el fin de determinar la presencia de la infección por H. pylori por ambos métodos. Finalmente se comparó el resultado de la anatomía patológica (patrón de oro) con el de TRU. Resultados: Se evaluó 181 pacientes, la edad promedio fue 52,8±13,5 años. La sensibilidad, especificidad, valor predictivo negativo (VPN), valor predictivo positivo (VPP) a los 20 minutos fueron de 86,8%, 98,5%, 81,5% y 99% y a las 24 horas 97,3%, 99,5%, 95,7% y 99,1% respectivamente. Conclusión: El TRU es un test confiable, accesible y de fácil aplicación para hacer el diagnóstico de la infección por H. pylori.


Objective: To validate a rapid urease test (RUT) in Cayetano Heredia Hospital (HCH) in Lima, Peru. Materials and methods: This is a prospective observational study that included 181 patients over 18 years old with dyspeptic symptoms. All of them underwent upper gastrointestinal endoscopy at the Department of Gastroenterology at HCH. They had not received, during the last four weeks, proton pump inhibitors (PPIs), bismuth or antibiotics. Two biopsies of antrum were taken, one to perform the TRU (Sensibacter pylori test®) and the other one for pathology, in order to determine by both methods the presence of H. pylori infection. TRU’s results were compared with pathology´s (gold standard). Results: 181 patients, average age 52.8±13.5 years, were evaluated. The sensitivity, specificity, negative predictive value (NPV), positive predictive value (PPV) at 20 minutes were 86.8%, 98.5%, 81.5% and 99% and at 24 hours 97.3%, 99.5%, 95.7% y 99.1% respectively. Conclusion: The rapid urease test is a reliable, accessible and easy to apply test for the diagnosis of H. pylori infection.


Assuntos
Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Urease/metabolismo , Helicobacter pylori/enzimologia , Infecções por Helicobacter/diagnóstico , Mucosa Gástrica/metabolismo , Peru , Biópsia , Biomarcadores/metabolismo , Valor Preditivo dos Testes , Estudos Prospectivos , Helicobacter pylori/isolamento & purificação , Infecções por Helicobacter/patologia , Sensibilidade e Especificidade , Mucosa Gástrica/microbiologia , Mucosa Gástrica/patologia , Hospitais
4.
The Korean Journal of Internal Medicine ; : 40-45, 2016.
Artigo em Inglês | WPRIM | ID: wpr-149376

RESUMO

BACKGROUND/AIMS: To evaluate a new monoclonal antibody for Helicobacter pylori urease in gastric tissue. METHODS: A total of 107 volunteers were enrolled. All subjects underwent a 13C-urea breath test and esophagogastroduodenoscopy. Gastric aspirates were analyzed for pH and ammonia. Six biopsy specimens in the gastric antrum and body were obtained for a rapid urease test and histology. The new monoclonal antibody-based H. pylori urease test (HPU) was performed to rapidly and qualitatively detect urease in two biopsy specimens. RESULTS: H. pylori infection was diagnosed in 73 subjects. The sensitivity and specificity of the HPU was 89% and 74%, respectively. The subjects were divided into two groups: one with true-positive and true-negative HPU results (n = 90) and the other with false-positive and false-negative HPU results (n = 17). Across all subjects, ammonia levels were 900.5 +/- 646.7 and 604.3 +/- 594.3 mumol/L (p > 0.05), and pH was 3.37 +/- 1.64 and 2.82 +/- 1.51 (p > 0.05). Sensitivity was higher in the presence of atrophic gastritis or intestinal metaplasia. CONCLUSIONS: HPU detected H. pylori in approximately 10 min. Gastric aspirate ammonia and pH levels did not affect the test results. Sensitivity was good in the presence of atrophic gastritis or intestinal metaplasia.


Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Anticorpos Monoclonais/imunologia , Proteínas de Bactérias/análise , Biomarcadores/análise , Biópsia , Reações Falso-Negativas , Reações Falso-Positivas , Gastrite Atrófica/diagnóstico , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/enzimologia , Testes Imunológicos , Metaplasia , Valor Preditivo dos Testes , Antro Pilórico/microbiologia , Reprodutibilidade dos Testes , Fatores de Tempo , Urease/análise , Fluxo de Trabalho
5.
GEN ; 66(2): 93-99, jun. 2012. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-664209

RESUMO

Existen múltiples métodos para diagnosticar la infección por H. pylori. Se plantea correlacionar la prueba del aliento con los hallazgos histopatológicos en el antro asociados a la infección por H. pylori en la población de Mucuchíes. Se incluyeron 108 pacientes entre 18 y 75 años. Se realizó prueba del aliento con 14C y endoscopia digestiva superior con obtención de cuatro muestras de mucosa antral. Se determinaron la sensibilidad, especificidad, valor predictivo positivo (VPP) y negativo (VPN), y la concordancia con la biopsia, además de correlaciones entre los valores de la prueba con la histopatología. Se obtuvieron 74 pacientes copositivos, con una sensibilidad del 87,05%, y una especificidad del 73,91% para la prueba del aliento. El VPP fue del 92,5%, y el VPN fue de 60,71%. El Kappa fue de 0,56. Se encontró correlación lineal entre la prueba del aliento, la densidad y la histología. La prueba del aliento constituye una herramienta útil en el diagnóstico de la infección por H. pylori y sus manifestaciones histopatológicas asociadas


There are many methods to diagnose H. pylori infection. It is proposed to compare the cuantitative14C Urea Breath Test with the histopathological features in the antrum related to H. pylori in the population of Mucuchíes. 108 patients between 18 and 75 years were included to perform the 14C Urea Breath Test, and the upper digestive endoscopy with four biopsy samples obtained from the gastric antrum. The sensitivity, specificity, PPV, NPV and Kappa index were determined for the 14C Urea Breath Test, and correlations between the 14C Urea Breath Test and histological features were determined too. We obtained 74 copositive patients, with a sensitivity of 87,05% and specificity of 73,91% for the 14C UBT. The PPV was of 92,5% and the NPV was of 60,71%. The Kappa index was of 0,56. A linear correlation between the test and the histological features was found. The 14C UBT is a very useful tool in the diagnosis of H. pylori infection and its related histopathological features


Assuntos
Feminino , Adulto Jovem , Protocolos Clínicos , Helicobacter pylori/enzimologia , Helicobacter pylori/isolamento & purificação , Infecções Bacterianas/diagnóstico , Bacteriologia , Gastroenterologia
6.
Indian J Med Microbiol ; 2011 Oct-Dec; 29(4): 379-382
Artigo em Inglês | IMSEAR | ID: sea-143860

RESUMO

Introduction: Presence of blood in the stomach has been thought to affect the performance of diagnostic tests used in detecting Helicobacter pylori (H. pylori) in the stomach. This study evaluated the effect of blood on the efficacy of rapid urease test (RUT) and microscopic appearance of the biopsy after staining with Giemsa stain. Materials and Methods: Patients with bleeding oesophageal varices who met the inclusion criteria were tested for H. pylori by RUT and microscopic examination of the biopsy. A repeat endoscopy, RUT and histology were done one month following initial presentation. The performance of the diagnostic tests was evaluated with and without the presence of intraluminal blood. A combined result of the two tests, RUT and histology, carried out in presence or absence of blood for the diagnosis of H. pylori, when considered together was considered as the gold standard. Results: Thirty six patients included in the study were in the ages ranging between 15-60 years (mean age = 44.14 years ±2.1). The combination of tests at both visits showed 20/36 (55.6%) patients were positive for H. pylori. The decrease in H. pylori positivity in the presence of blood was significant for RUT (8.3% vs. 38.9%; P=0.005) and combined test (19.4% vs. 47.2%; P=0.02) but the decrease in positivity for histology (11.1% vs 30.6%) was not significant (P=0.08). In the presence of blood, the sensitivity of RUT, histology and combined tests were 15%, 20% and 35%, respectively. In the absence of blood, the sensitivity of RUT, histology and combination of tests was 70%, 55% and 85%, respectively. Conclusion: Blood in the stomach significantly decreased the sensitivity of RUT, histology and the combination of both. Negative results of these tests in acute upper gastro intestinal (GI) bleeding should therefore be interpreted carefully.


Assuntos
Adolescente , Adulto , Biópsia , Sangue , Endoscopia Gastrointestinal/métodos , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/citologia , Helicobacter pylori/enzimologia , Helicobacter pylori/isolamento & purificação , Histocitoquímica/métodos , Humanos , Masculino , Microscopia/métodos , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Estômago/microbiologia , Estômago/patologia , Urease/análise , Adulto Jovem
8.
Yonsei Medical Journal ; : 39-44, 2010.
Artigo em Inglês | WPRIM | ID: wpr-39512

RESUMO

PURPOSE: The role of the Ferric Uptake Regulator (FUR) in the acid resistance of Helicobacter pylori (H. pylori) has been thought to be independent of urease. However, we demonstrated in this study that Fur influences urease activity. MATERIALS AND METHODS: A fur knockout mutant of H. pylori was constructed by replacing the Fur gene with a kanamycin resistant marker gene. The wild-type H. pylori and fur mutant were compared for survival. The integrity of the inner membrane of the bacteria was evaluated by confocal microscopy using membrane-permeant and -impermeant fluorescent DNA probes. Urease activity of intact H. pylori was measured between pH 3 and 8. Real time PCR of both strains was performed for urease genes including ureI, ureE, ureF, ureG, and ureH. RESULTS: The fur deletion affected the survival of H. pylori at pH 4. The urease activity curve of the intact fur mutant showed the same shape as the wild-type but was 3-fold lower than the wild-type at a pH of less than 5. Real time PCR revealed that the expression of all genes was consistently down-regulated in the fur mutant. CONCLUSION: The results of this study showed that fur appears to be involved in acid resistant H. pylori urease activity.


Assuntos
Proteínas de Bactérias/genética , Helicobacter pylori/enzimologia , Concentração de Íons de Hidrogênio , Microscopia Confocal , Modelos Biológicos , Mutação , Proteínas Repressoras/genética , Urease/metabolismo
9.
Artigo em Inglês | IMSEAR | ID: sea-139715

RESUMO

The oral cavity has been considered a potential reservoir for Helicobacter pylori (H pylori) , from where the organism causes recurrent gastric infections. Aim: With this case-control study we tried to evaluate the role of H pylori in the etiology of mucosal inflammation, a condition that compounds the morbid state associated with oral submucous fibrosis (OSF). Materials and Methods : Subjects ( n = 150) were selected following institutional regulations on sample collection and grouped into test cases and positive and negative controls based on the presence of mucosal fibrosis and inflammation. The negative controls had none of the clinical signs. All patients underwent an oral examination as well as tests to assess oral hygiene/periodontal disease status; a rapid urease test (RUT) of plaque samples was also done to estimate the H pylori bacterial load. We used univariate and mutivariate logistic regression for statistical analysis of the data and calculated the odds ratios to assess the risk posed by the different variables. Results : The RUT results differed significantly between the groups, reflecting the variations in the bacterial loads in each category. The test was positive in 52% in the positive controls (where nonspecific inflammation of oral mucosa was seen unassociated with fibrosis), in 46% of the test cases, and in 18% of the negative controls (healthy volunteers) (χ2 = 13.887; P < 0.01). A positive correlation was seen between the oral hygiene/periodontal disease indices and RUT reactivity in all the three groups. Conclusions: The contribution of the H pylori in dental plaque to mucosal inflammation and periodontal disease was significant. Logistic regression analysis showed gastrointestinal disease and poor oral hygiene as being the greatest risk factors for bacterial colonization, irrespective of the subject groups. A positive correlation exists between RUT reactivity and the frequency of mucosal inflammation.


Assuntos
Estudos de Casos e Controles , Contagem de Colônia Microbiana , Cárie Dentária/complicações , Placa Dentária/microbiologia , Complicações do Diabetes , Dispepsia/microbiologia , Feminino , Glossite/microbiologia , Infecções por Helicobacter/complicações , Helicobacter pylori/enzimologia , Helicobacter pylori/isolamento & purificação , Humanos , Hipertensão/complicações , Masculino , Índice de Higiene Oral , Fibrose Oral Submucosa/microbiologia , Perda da Inserção Periodontal/microbiologia , Índice Periodontal , Bolsa Periodontal/microbiologia , Estomatite/microbiologia , Urease/análise
11.
Braz. j. med. biol. res ; 39(7): 851-861, July 2006. tab, graf
Artigo em Inglês | LILACS | ID: lil-431568

RESUMO

Ureases are enzymes from plants, fungi and bacteria that catalyze the hydrolysis of urea to form ammonia and carbon dioxide. While fungal and plant ureases are homo-oligomers of 90-kDa subunits, bacterial ureases are multimers of two or three subunit complexes. We showed that some isoforms of jack bean urease, canatoxin and the classical urease, bind to glycoconjugates and induce platelet aggregation. Canatoxin also promotes release of histamine from mast cells, insulin from pancreatic cells and neurotransmitters from brain synaptosomes. In vivo it induces rat paw edema and neutrophil chemotaxis. These effects are independent of ureolytic activity and require activation of eicosanoid metabolism and calcium channels. Helicobacter pylori, a Gram-negative bacterium that colonizes the human stomach mucosa, causes gastric ulcers and cancer by a mechanism that is not understood. H. pylori produces factors that damage gastric epithelial cells, such as the vacuolating cytotoxin VacA, the cytotoxin-associated protein CagA, and a urease (up to 10 percent of bacterial protein) that neutralizes the acidic medium permitting its survival in the stomach. H. pylori whole cells or extracts of its water-soluble proteins promote inflammation, activate neutrophils and induce the release of cytokines. In this paper we review data from the literature suggesting that H. pylori urease displays many of the biological activities observed for jack bean ureases and show that bacterial ureases have a secretagogue effect modulated by eicosanoid metabolites through lipoxygenase pathways. These findings could be relevant to the elucidation of the role of urease in the pathogenesis of the gastrointestinal disease caused by H. pylori.


Assuntos
Humanos , Animais , Canavalia/enzimologia , Eicosanoides/metabolismo , Infecções por Helicobacter/microbiologia , Helicobacter pylori/enzimologia , Proteínas de Plantas/biossíntese , Urease/biossíntese , Relação Dose-Resposta a Droga , Duodenopatias/metabolismo , Duodenopatias/microbiologia , Infecções por Helicobacter/metabolismo , Gastropatias/metabolismo , Gastropatias/microbiologia
12.
Southeast Asian J Trop Med Public Health ; 2006 Mar; 37(2): 312-9
Artigo em Inglês | IMSEAR | ID: sea-34144

RESUMO

We developed an in-house rapid urease test (iRUT) and evaluated the efficacy and the agreement of the iRUT and the cRUT compared with culture and histology for the detection of H. pylori infection. Five iRUT media were tested with H. pylori isolates and other bacteria. The most suitable iRUT medium was further evaluated for detection of H. pylori infection. Gastric biopsies from 120 patients were diagnosed by culture, iRUT, cRUT and histology. The results of the iRUT and cRUT were read at 30 minutes, 1 hour and up to 24 hours. A true positive result was either the culture or both the RUT (cRUT or iRUT) and the histological examination being positive. The sensitivity and specificity of the iRUT result at 30 minutes, 1 hour and up to 24 hours were 77.1% and 100%, 77.6% and 100%, and 94.1% and 94.2%, respectively. Values for the same parameters of cRUT were 87.5% and 100%, 89.8% and 100%, and 100% and 94.2%, respectively. The agreement between the iRUT and cRUT was very good (kappa values > or = 0.82). Our results indicate that the iRUT is a-sensitive, specific and cost effective test. It can be appropriately applied for detecting H. pylori infection in gastric biopsy specimens.


Assuntos
Biópsia , Contagem de Colônia Microbiana , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/enzimologia , Humanos , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Estômago/microbiologia , Fatores de Tempo , Urease/metabolismo
13.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 22-4, 2004.
Artigo em Inglês | WPRIM | ID: wpr-634068

RESUMO

The gene encoding urease subunit A (ureA) of Helicobacter pylori (H. pylori) was cloned from H. pylori isolate by polymerase chain reaction (PCR). Sterile distilled water instead of DNA served as negative control. The nucleotide sequence of the amplified product was determined. Homologous analysis of the ureA against that reported by Clayton CL and the GenBank and SwissProt databases were performed with the BLAST program at the Genome Net through the Internet. 0.8 kb PCR product was amplified from all H. pylori clinical isolators. The nucleotide sequence of the ureA was determined. The nucleotide sequence of the ureA began with ATG as the initiation codon and terminated in TAA as stop codon. The coding regions had a 44% G + C content. The DNA sequence was 98% homologous to that reported by Clayton CL (688 out of 702 residues were identical). The derived amino-acid sequences of the ureA were 99% homologous to that reported by Clayton CL (232 out of 234 residues were identical). The nucleotide sequence and the predicted protein showed significant homology to ureA of H. pylori in the NCBI Entrez database.


Assuntos
Sequência de Bases , Clonagem Molecular , DNA Bacteriano/química , DNA Bacteriano/genética , Genes Bacterianos , Código Genético , Infecções por Helicobacter/microbiologia , Helicobacter pylori/enzimologia , Helicobacter pylori/genética , Helicobacter pylori/isolamento & purificação , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Transcrição Gênica , Urease/genética , Urease/metabolismo
14.
Indian J Pathol Microbiol ; 2003 Jul; 46(3): 511-4
Artigo em Inglês | IMSEAR | ID: sea-73817

RESUMO

Helicobacter pylori is one of the most common chronic bacterial infection in humans linked to acid peptic diseases, gastric carcinomas and lymphomas. The bacilli produces large amounts of urease and this property has formed the basis of detection of H. pylori by the Christensen's urease test. Where endoscopy is not clinically indicated, serology may be used to establish the diagnosis. This study was undertaken to diagnose H. pylori with the help of Christensen's urease test on endoscopic biopsy specimens & correlated with the detection in Sera, of IgG antibodies against H. pylori, by ELISA technique. The study was conducted on 100 patients suffering from acid peptic disorders out of which 40 (40%) tested positive for H. pylori both by urease and serology. Christensen's urease and ELISA were found to have sensitivities of 85.7% & 90.9% and specificities of 96% and 87.5% respectively. Christensen's urease was taken as a standard method of diagnosis and its correlation with ELISA worked out to (+1) which meant there was a strong positive association between both the tests. Hence either test could be used for primary diagnosis of H. pylori instead of histopathological study and/or culture of H. pylori.


Assuntos
Anticorpos Antibacterianos/sangue , Dispepsia/microbiologia , Ensaio de Imunoadsorção Enzimática , Esofagite/microbiologia , Feminino , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/enzimologia , Humanos , Masculino , Úlcera Péptica/microbiologia , Urease/análise
15.
Acta gastroenterol. latinoam ; 33(2): 73-76, 2003. ilus, tab
Artigo em Português | LILACS | ID: lil-420385

RESUMO

The aim of our study was to develop a rapid diagnostic urease test to demonstrate the presence of Helicobacter pylori in the Endoscopy room. MATERIALS AND METHODS: 200 consecutive patients referred to gastroscopy for different indications, were included in this study. One antral biopsy sample was obtained to be immersed in our test. The same sample was used for histological evaluation, considered to be the gold standard method for diagnose of Helicobacter pylori infection. RESULTS: 135 patients (67.5%) were found positives and 65 patients (32.5%) were negatives in our test. 128 patients (64%) showed Helicobacter pylori on histological examination. Our test showed a sensitivity of 91%, specificity of 88.1%, and positive and negative predictive values of 95% and 80% respectively. A remarkable correlation between density of Helicobacter pylori and reading time was also observed, where a high density of the bacteria reduced the reaction time in this liquid test. Furthermore, an overall accuracy of 90% was shown, which is comparable with other available commercial tests. CONCLUSION: LUT is easy to handle, cost effective and fast, with a high positive predictive value.


Assuntos
Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Ensaios Enzimáticos Clínicos , Mucosa Gástrica/microbiologia , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/isolamento & purificação , Urease/análise , Ensaios Enzimáticos Clínicos , Endoscopia Gastrointestinal , Mucosa Gástrica/patologia , Helicobacter pylori/enzimologia , Estudos Prospectivos , Antro Pilórico/microbiologia , Antro Pilórico/patologia , Sensibilidade e Especificidade , Urease/metabolismo
16.
Rev. gastroenterol. Méx ; 63(3): 135-42, jul.-sept. 1998. tab, ilus
Artigo em Espanhol | LILACS | ID: lil-240904

RESUMO

Existen varios métodos para la detección de la infección por Helicobacter pylori, algunos de ellos (invasivos) requieren de un procedimiento endoscópico y biopsia como la prueba rápida de ureasa, el cultivo y la histología. Recientemente se han desarrollado y aprobado métodos no invasivos, sensibles, específicos, fáciles de realizar y bien aceptados por los pacientes como la prueba de aliento, basada en la hidrólisis de la urea marcada por la ureasa del Helicobacter pylori formando amonio y bicarbonato. El CO2 marcado pasa, a través de la circulación, a los pulmones y es espirado en el aliento de donde puede ser recolectado. La urea puede ser marcada en su átomo de carbono existiendo dos opciones: 13C estable, no radioactivo y 14 C inestable y radioactivo. La prueba de aliento con 13C se basa en la diferencia de masa atómica entre el 12C y 13C y requiere para su cuantificación de un espectómetro de masas y aproximadamente 40 minutos para su realización. La prueba de aliento con 14C contiene 1 uCi (un micro-curie) de radioactividad que corresponde a 1/300 de la radiación recibida en un año del medio ambiente; su realización toma 10 minutos y la lectura se realiza en un contador de centello. Ambas pruebas, no invasivas, han demostrado sensibilidad y especificidad comparables con los ®estándares de oro¼ establecido para el diagnóstico de infección por Helicobacter pylori


Assuntos
Humanos , Testes Respiratórios , Isótopos de Carbono , Radioisótopos de Carbono , Reações Falso-Negativas , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/enzimologia , Sensibilidade e Especificidade , Urease/análise , Ureia/metabolismo
17.
Indian J Pathol Microbiol ; 1995 Oct; 38(4): 349-54
Artigo em Inglês | IMSEAR | ID: sea-75705

RESUMO

A modified Rapid urease test developed by us was evaluated as a screening test for Helicobacter pylori (H. pylori) during and endoscopy survey on patients with Acid Peptic Diseases (APD) and Non Ulcer Dyspepsia (NUD). This was compared with commercially available CLO (Campylobacter Like Organism) test, culture and histopathological examination. The modified Rapid urease test gave a sensitivity of 89.83% and a specificity of 100%, when compared to 95% sensitivity and specificity for commercially available CLO test. Our modified Rapid urease test is simple, economical and a quick test in identifying H. pylori in routine screening of patients with APD and NUD.


Assuntos
Adolescente , Adulto , Idoso , Técnicas Bacteriológicas , Feminino , Mucosa Gástrica/microbiologia , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/enzimologia , Humanos , Masculino , Pessoa de Meia-Idade , Gastropatias/diagnóstico , Urease/análise
18.
GEN ; 49(3): 208-11, jul.-sept. 1995. tab
Artigo em Espanhol | LILACS | ID: lil-163470

RESUMO

Actualmente se encuentran disponibles un gran número de ensayos inmunoenzimáticos para la determminación de la respuesta humoral generada durante la infección por Helicobacter Pylori. Sin embargo, no existe un concenso sobre el método ideal a utilizar para evaluar la infección debida a este microorganismo. En el presente estudio se evaluó la sensibilidad de un ensayo inmunoenzimático desarrollado en el laboratorio, empleando 38 sueros de pacientes con úlcera duodenal y cultivo positivo para H. pylori y 41 sueros de niños sanos menores de 10 años. En el ensayo se utilizaron extractos proteícos obtenidos de la sonicación de cinco cepas aisladas de los pacientes con úlcera duodenal. Los niveles de la IgG del grupo de pacientes fueron significativamente mayores (p < 0.001) que en el grupo control. El límite de positividad de 0.231 fue determinado empleando la media + 3ds, de los títulos obtenidos en el grupo control. Los resultados del ensayo inmunoenzimático mostraron que una dilución de 1:300 de los sueros y una concentración de antígeno de 5 ug/ml sensibilidad del 92 por ciento en la determinación de la infección por H. pylori


Assuntos
Humanos , Ensaio de Imunoadsorção Enzimática/estatística & dados numéricos , Helicobacter pylori/enzimologia , Helicobacter pylori/patogenicidade , Imunoglobulina M/análise , Imunoglobulina M/uso terapêutico , Úlcera Duodenal/diagnóstico , Úlcera Duodenal/enzimologia
19.
Artigo em Inglês | IMSEAR | ID: sea-43829

RESUMO

The prevalence of H.pylori in Thailand is high compared with Western countries and is the same as in China. We suggest either rapid urease test (CLO test) or Giemsa stain to be a rapid, reliable and convenient detection method for H.pylori and is also suitable for use in follow-up studies by gastroenterologists.


Assuntos
Adolescente , Adulto , Idoso , Dispepsia/microbiologia , Feminino , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/enzimologia , Humanos , Masculino , Pessoa de Meia-Idade , Coloração e Rotulagem , Urease/metabolismo
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