Molecular technique for detection and identification of Helicobacter pylori in clinical specimens: a comparison with the classical diagnostic method

Nevoa, Jessica Caraiola Nevoa Caraiola; Rodrigues, Roger Luiz; Menezes, Gabriela L.; Lopes, Andressa Rodrigues; Nascimento, Hemelly Faria Nascimento Faria; Santiago, Silvana Barbosa; Morelli, Marcos L.; Barbosa, Mônica Santiago

doi:10.5935/1676-2444.20170003

Cited by 13 publications

(26 citation statements)

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“…Our results were comparable with Parihar et al [44] and higher than Allahverdiyev et al [45]. Nevoa et al [46] found that the rate in the detection of H. pylori by the molecular method was significantly higher when compared to the rapid urease test.…”

supporting

confidence: 88%

Helicobacter Pylori Infection in Egyptian Patients With Dyspepsia: Diagnostic, Demographic, Endoscopic and Clinical Characteristics.

Diab¹,

Shenawy²,

Shemis³

et al. 2018

IJAR

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Helicobacter pylori, PCR, rapid urease test, stool antigen. was revealed in 27(45%) and 10 (16.7%) had peptic ulcer disease (PUD). Conclusions: H. pylori infection rate in Egyptian patients with dyspepsia was high and gastritis was the most revealed finding upon endoscopy. No risk factors were associated with H. pylori infection among the studied adult patients. Combined rapid urease and stool antigen tests can be relied upon for detecting H. pylori infection.

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supporting

confidence: 88%

Helicobacter Pylori Infection in Egyptian Patients With Dyspepsia: Diagnostic, Demographic, Endoscopic and Clinical Characteristics.

Diab¹,

Shenawy²,

Shemis³

et al. 2018

IJAR

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“…The developed RUT should be read within 24 h with a closed lid to avoid false-positive results from contamination by bacteria that are not H. pylori , such as Proteus , Yersinia , Klebsiella , and Pseudomonas , which can produce urease; urea is hydrolyzed to produce ammonia, leading to an increase in pH causing a positive test result [ 26 ]. In this study, flaws were found in the substance used as it was not only opaque white plastic, but also too thick, making it difficult to observe any color change.…”

Section: Discussionmentioning

confidence: 99%

Development of an easy-to-use urease kit for detecting Helicobacter pylori in canine gastric mucosa

et al. 2021

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Background and Aim: Helicobacter pylori is an important pathogen in humans and animals involved in chronic gastritis, leading to the development of gastric cancer. Urease produced by H. pylori is an enzyme that promotes bacterial colonization and can be used clinically as a biomarker of H. pylori infection as part of a rapid urease test (RUT). A test with high specificity (95-100%) would be more convenient and faster than histopathology, bacterial culture, and polymerase chain reaction (PCR). The aim of this study was to develop a simple, cheap, and fast kit for detecting H. pylori infection in the gastric mucosa of canines, which can be used in clinical practice for diagnosing infection with this bacterium. Materials and Methods: The RUT assays developed were prepared using 1% agar, 1% sodium phosphate monobasic, and 1% urea followed by the addition of 3% methyl red indicator. The cutoff value of sensitivity of the RUT assay was established using the urease of H. pylori ATCC 43504 and color change was monitored for 24 h. Comparisons of the sensitivity to H. pylori ATCC 43504 were made between the developed RUT assays and the Hp Fast™ commercial kit. Then, the limit of detection for H. pylori ATCC 43504 number was analyzed by the SYBR Green real-time PCR assay to measure the copy number of the ureC gene. Gastric biopsy samples from the antrum, body, and fundus of the stomach were collected from eight canines presenting with vomiting and gastroenteritis. Analyses were performed on fresh samples using the developed RUT assays and the Hp Fast™ commercial kit, which were read within 24 h; then, the results were confirmed with SYBR Green real-time PCR. The specificity of the RUT assays was tested with a number of different bacteria, including Staphylococcus pseudintermedius, Proteus spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterococcus spp., Escherichia coli, and Salmonella spp.; H. pylori ATCC 43504 was used as a positive control. Results: The results showed that the developed assays were sensitive to the urease enzyme at 0.1 mg/mL. The lowest detection limit of this assay for H. pylori ATCC 43504 was found to be 102 copies at 30 min. The sensitivity of detection of H. pylori in gastric biopsies of canines occurred in a minimum of 30 min. The RUT showed similar results to the Hp Fast™ commercial kit. In the developed RUT, the color change of the test from red to yellow could be clearly distinguished between the color of the positive test and the negative one; however, in the commercial Hp Fast™, it was difficult to observe the gel color change in the negative pH range of 5.8 and the positive pH of 6.5. The developed RUT was specific for H. pylori and did not detect any of the other tested bacteria. The test kit can also be stored for 6 months at 4°C. Conclusion: The sensitivity of the developed assays allowed the detection of urease enzyme at a minimum concentration of 0.1 mg/mL. Our RUT could also detect H. pylori from one in eight canine specimens at a minimum of 102 copies within 30 min. This RUT is specific to H. pylori as it did not detect any of the other tested bacteria.

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“…' O H.pylori é uma bactéria microaerófila gram-negativa que infecta as células epiteliais que secretam muco no estômago humano. Essa bactéria pode propiciar o surgimento de diversas doenças gástricas, como gastrite, úlceras pépticas e câncer gástrico 1 . Na atualidade, são comprovados três meios de transmissão: fecal-oral, oral-oral e iatrogênica.…”

Section: Medicinaunclassified

Métodos diagnósticos para detecção da infecção pelo h.pylori

Vargas¹,

Deprá²,

Nascimento³

et al. 2022

PRMJ

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MedicinaCopyright Vargas et al. Este é um artigo publicado em acesso aberto (Open Access) sob a licença Creative Commons Attribution, que permite uso, distribuição e reprodução em qualquer meio, sem restrições desde que o trabalho original seja corretamente citado. Trabalho realizado na Faculdade Metropolitana da Amazônia -FAMAZ, Belém, PA, Brasil.resumo Objetivo: Realizar uma revisão sistemática dos métodos diagnósticos para detecção da infecção pelo H.pylori. Método: Revisão sistemática de artigos originais que analisam os métodos diagnósticos para a detecção da infecção pelo H.pylori, no período de 2014 até 2018 utilizando o Google Acadêmico, Bireme, Scielo e LILACS como base de dados. Resultados: Diversos métodos diagnósticos são utilizados para detecção do H.pylori, classificados em invasivos e não invasivos. Dentre os métodos invasivos, o mais indicado é o teste molecular (PCR) e dentre os métodos não invasivos, o teste respiratório com 13C-uréia é o com maior especificidade e sensibilidade. Conclusão: Não existe nenhum teste considerado padrão-ouro, fazendo-se necessário analisar cada situação específica e buscar a melhor alternativa diante da condição clínica do paciente, assim como a disponibilidade do exame nos diversos serviços especializados, contribuindo assim, para um diagnóstico seguro, preciso e eficaz. Descritores: helicobacter pylori; diagnóstico; métodos e infecção.summary Purpose: Perform a systematic review of the diagnostic methods for detecting infection by H.pylori. Methods: The method was a systematic review of original articles that analyze the diagnostic methods for the detection of H. pylori infection from 2015 to 2018 using Google Scholar, Bireme, Scielo and LILACS as a database. Results: Several diagnostic methods are used to detect H. pylori, classified as invasive and noninvasive. Among the invasive methods, the molecular test (PCR) is the most indicated and among the noninvasive methods, the respiratory test with 13C-urea is the one with the highest specifity and sensivity. Conclusion: There is no gold standard test, making it necessary to analyze each specific situation and seek the best alternative in view of the clinical condition of the patient, as well as the availability of the examination in the various specialized services, thus contributing to a safe, accurate and effective. informação sobre os autores LJV, LRD e LEMCN são acadêmicos do curso de medicina da Faculdade Metropolitana da Amazônia -FAMAZ. APSOB, HCRG e RBM são professores orientadores da Faculdade Metropolitana da Amazônia -FAMAZ. Contribuição dos autores LJV, LRD, LEMCN contribuíram escrevendo, editando e revisando o manuscrito com base na luz da literatura atual, e seguindo as coordenadas de seus orientadores; APSOB, HCRG e RBM, seguiram orientando e editando os recortes textuais e pontuando os principais pontos críticos relevantes para a construção e elaboração textual. Todos os autores leram e aprovaram a versão final submetida ao Pará Research Medical Journal.

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Molecular technique for detection and identification of Helicobacter pylori in clinical specimens: a comparison with the classical diagnostic method

Cited by 13 publications

References 0 publications

Helicobacter Pylori Infection in Egyptian Patients With Dyspepsia: Diagnostic, Demographic, Endoscopic and Clinical Characteristics.

Helicobacter Pylori Infection in Egyptian Patients With Dyspepsia: Diagnostic, Demographic, Endoscopic and Clinical Characteristics.

Development of an easy-to-use urease kit for detecting Helicobacter pylori in canine gastric mucosa

Métodos diagnósticos para detecção da infecção pelo h.pylori

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