Non-invasive Diagnostic of Helicobacter pylori in Stools by Nested-qPCR

The aim of this study was to develop a non-invasive diagnostic test for the detection of Helicobacter pylori in stool samples from digestive symptomatic patients, using a new protocol of nested-qPCR. A total of 143 patients were invited to participate in the study. A gastric biopsy of each patient was collected for Rapid Urease Testing (RUT) and histology by Giemsa stain. A fecal sample for nested-qPCR analysis was also obtained. DNA was extracted from the fecal samples, and conventional PCR followed by qPCR of the ureC gene of H. pylori was carried out. We evaluated the presence of H. pylori, in 103 females and 40 males, mean (± SD) age of 56.5 ± 14.18. The sensitivity of RUT to detect the infection was 67.0% (95% C.I.: 57.2 – 75.8) and specificity was 92.3% (95% C.I.: 76.5 – 99.1). Histology by Giemsa stain, commonly used as a reference for H. pylori detection, showed a sensitivity of 98.6% (95% C.I.: 92.5 – 100.0) and a specificity of 89.7% (95% C.I.: 72.7 – 97.8). In contrast, detection of H. pylori infection in stools by nested-qPCR showed a sensitivity of 100% (95% C.I.: 94.9 – 100.0) and a specificity of 83.9% (95% C.I.: 66.3 – 94.6). Our test, based in nested-qPCR is a better diagnostic alternative than conventional RUT, and is similar to histology by Giemsa stain in the detection of H. pylori, by which the test could be used for non-invasive diagnosis in clinical practice.

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Detection of Helicobacter pylori in gastric biopsy and resection specimens

Vojnosanitetski pregled ◽

10.2298/vsp0501039b ◽

2005 ◽

Vol 62 (1) ◽

pp. 39-43 ◽

Cited By ~ 1

Author(s):

Tatjana Babic ◽

Hakija Basic ◽

Biljana Selimovic-Miljkovic ◽

Branislava Kocic ◽

Gordana Tasic

Keyword(s):

Duodenal Ulcer ◽

Alkaline Phosphatase ◽

Helicobacter Pylori ◽

Gastric Biopsy ◽

Giemsa Stain ◽

Biopsy Specimens ◽

H Pylori ◽

Antral Biopsy ◽

Better Than ◽

Immunohistochemical Identification

Aim. To compare the sensitivity of detecting H. pylori in gastric biopsy and resection specimens using modified Giemsa stain and immunohistochemistry, using a commercially available anti-H. pylori antibody (Dako, Denmark). Methods. Gastric antral biopsy specimens showing chronic gastritis (28 cases) together with tissue blocks from gastrectomy specimens for duodenal ulcer (2 cases) were stained with modified Giemsa and immunoenzymatic alkaline phosphatase - anti-alkaline phosphatase (APAAP) method, and were carefully examined for the presence of H. pylori. Results. Using a modified Giemsa stain, the spiral shaped bacteria of H. pylori stained blue, were attached to the brush border of the gastric foveolar epithelial cells. However, the specificity of modified Giemsa stain depended on the morphological appearance of H. pylori. The specificity of immunostaining permitted detection of low numbers or even single organisms. In all cases bacteria were more prominent and easier to detect in immunostained preparations. H. pylori was identified in 22 (73.3%) of 30 sections stained with modified Giemsa stain, but it could be identified with greater frequency in sections stained with APAAP, in 27 (90%) of 30 sections. Conclusion. Immunohistochemical identification of H. pylori was better than Giemsa stain for detecting that organism.

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Helicobacter pylori prevalence in healthy Mexican children: comparison between two non-invasive methods

PeerJ ◽

10.7717/peerj.11546 ◽

2021 ◽

Vol 9 ◽

pp. e11546

Author(s):

Verónica I. Martínez-Santos ◽

Manuel Hernández Catalán ◽

Luis Octavio Ojeda Salazar ◽

Octavio Andrei Orozco Gómez ◽

Sandra Ines Lorenzo ◽

...

Keyword(s):

Helicobacter Pylori ◽

Dental Plaque ◽

Rrna Gene ◽

Suspected Infection ◽

Stool Antigen Test ◽

Non Invasive ◽

Asymptomatic Children ◽

Feasible Alternative ◽

Stool Samples ◽

H Pylori

Background Helicobacter pylori detection in asymptomatic children with suspected infection or with symptoms that suggest gastric pathology is problematic, since most of the methods depend on the endoscopic study, an invasive and expensive method. Non-invasive methods can be a feasible alternative but must be validated. The purpose of this study was to evaluate the concordance between H. pylori DNA detection in saliva and dental plaque by PCR, with antigen detection in stool by immunochromatography, among asymptomatic children in the state of Guerrero, Mexico. Methods Dental plaque, saliva, and stool samples were obtained from 171 children between 6 and 12 years old. H. pylori detection in saliva and dental plaque was performed by PCR using specific primers for the 16S rRNA gene, while the detection in stool samples was performed by immunochromatography using the CerTest kit. Results We found an overall H. pylori prevalence of 59.6% (102/171). Of the H. pylori positive children 18% (20/111) were positive in saliva samples, 28.1% (34/121) in dental plaque samples, and 50.4% (71/141) in stool samples. A higher prevalence was found in girls (64.7%, p = 0.002). Although some of the children declared some dyspeptic symptoms, these were no related to H. pylori. In conclusion, we found a high prevalence of H. pylori in asymptomatic children and the highest proportion was detected by stool antigen test, which was the most feasible method to detect H. pylori infection.

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Evaluation of Rapid stool antigen test for the diagnosis of Helicobacter pylori infection in patients with dyspepsia

IMC Journal of Medical Science ◽

10.3329/imcjms.v10i2.31108 ◽

2017 ◽

Vol 10 (2) ◽

pp. 39-44

Author(s):

Salma Khatun ◽

Fahmida Rahman ◽

Khandaker Shadia ◽

Indrajit Kumar Dutta ◽

Mohammad Nazmul Hoq ◽

...

Keyword(s):

Helicobacter Pylori ◽

Peptic Ulcer ◽

Helicobacter Pylori Infection ◽

Adult Patients ◽

Antigen Test ◽

Stool Antigen Test ◽

Non Invasive ◽

Stool Antigen ◽

Stool Samples ◽

H Pylori

Background and objectives: Several diagnostic assays are used for the detection of Helicobacter pylori infection in suspected peptic ulcer cases. H. pylori stool antigen test is a non-invasive method for the detection of active infection. The present study has evaluated the efficacy of rapid stool antigen test to diagnose H. pylori infection in patients with dyspepsia.Materials and methods: Adult patients with complains of dyspepsia attending the Department of Gastroenterology, Hepatobiliary and Pancreatic Diseases (GHPD) of BIRDEM hospital for endoscopy were included. Gastric biopsy, blood and stool samples were obtained from each participant after informed written consent. Rapid urease test (RUT), serum H. pylori immunoglobulin A (IgA) and IgG and rapid H. pylori stool antigen (HpSAg) tests were performed. Only stool samples were obtained from 31 neonates aged 1 to 30 days as negative control for HpSAg test.Results: A total of 91 adult patients with complain of dyspepsia were included in the study. Out of 91 cases, 17 (18.7%) and 74 (81.3%) had peptic ulcer and erosion respectively. HpSAg was positive in 63.7% cases compared to 42.9% and 62.6% respectively by RUT and IgA. The rate of HpSAg positivity was significantly higher (p<0.05) in ulcer compared to erosion cases. HpSAg test was positive in all (100%) RUT positive cases. Combination of HpSAg test and IgA yielded highest positive result in both ulcer (82.4%) and erosion (84%) cases. H. pylori IgG was positive in all cases.Conclusion: The study has demonstrated that HpSAg test is an effective and non-invasive diagnostic tool to detect active H. pylori infection in suspected dyspeptic patients.IMC J Med Sci 2016; 10(2): 39-44

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Perfil Epidemiológico de Pacientes Submetidos à Biópsia Gástrica em um Hospital Escola do Sul de Minas Gerais/ Epidemiological Profile of Patients Undergo Gastric Biopsy in a School Hospital of Minas Gerais

REVISTA CIÊNCIAS EM SAÚDE ◽

10.21876/rcsfmit.v4i3.229 ◽

1970 ◽

Vol 4 (3) ◽

pp. 48-57

Author(s):

Isabela Maria A. Ribeiro Simões ◽

Ana Carolina Mauad Coli ◽

Roseane de Souza Candido Irulegui

Keyword(s):

Helicobacter Pylori ◽

Age Groups ◽

Epidemiological Data ◽

Minas Gerais ◽

Gastric Biopsy ◽

Gastric Ulcers ◽

Histopathological Diagnosis ◽

Gastric Diseases ◽

H Pylori ◽

Epidemiological Profile

Objetivo: Determinar a prevalência de lesões benignas e neoplasia gástrica através do estudo de biópsias realizadas em um Hospital Escola do Sul de Minas Gerais, no período entre 2007 e 2011. Materiais e Métodos: A pesquisa documental foi quantitativa e retrospectiva, baseada na análise dos registros de biópsias e prontuários. Realizou-se o levantamento de dados referentes à idade, gênero, cor, profissão, diagnóstico histopatológico e presença de Helicobacter pylori nas amostras. Resultados: O número total de biópsias gástricas analisadas foi de 1225, cujo perfil populacional encontrado foi: idade média de 56,75 anos, sexo masculino (52%), cor branca (81,9 %), aposentado (30%). Os diagnósticos mais frequentes foram: gastrites (71,9%), pólipos (14,2%), adenocarcinomas (5,9%), úlceras gástricas (6%), linfomas (0,4%), sem alterações (0,4%) e outros (1,2%). Em outros, encontram-se achados de malignidade, metaplasia e xantelasma gástrico. Em relação à presença de Helicobacter pylori nas amostras, o resultado encontrado foi de24% positivas, 46% negativas e 30% não pesquisadas. Conclusão: Os resultados confirmam a alta frequência das doenças gástricas e sua incidência nas diversas faixas etárias, além do envolvimento do H. pylori em tais afecções. É de grande importância a caracterização dos dados epidemiológicos, o que permite prováveis direcionamentos para programas de prevenção e informação para a população. Palavras-chave: biópsia gástrica, gastropatia, perfil epidemiológico. ABSTRACTObjective: To determine the prevalence of benign lesions and gastric cancer through study of biopsies performed at a school hospital in southern Minas Gerais, in the period between 2007 and 2011.Materials and Methods: The research was quantitative and retrospective, based on analysis of biopsies records and medical records. We conducted the survey data regarding age, sex, color, profession, histopathological diagnosis and the presence of Helicobacter pylori in the samples. Results: The total number of gastric biopsies analyzed was 1225. Population listing was found: mean age of 56.75 years, male (52%), white (81.9%), retired (30%). The most frequent diagnoses were gastritis (71.9%), polyps (14.2%), adenocarcinomas (5.9%), gastric ulcers (6%), lymphoma (0.4%), unchanged (0, 4%) and others (1.2%). In others, there are: findings of malignancy, metaplasia, gastric xanthelasma. Regarding the presence of Helicobacter pylori in the sample, the result was: 24% positive, 46% negative, 30% non searched. Conclusion: The results confirm the high frequency of gastric diseases and their incidence in the various age groups additionally to the involvement of H. pylori in such conditions. It is of great importance to characterize the epidemiological data, allowing probable directions for prevention and information programs for population. Keywords: gastric biopsy, gastropathy, epidemiological profile

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Evaluation of Better Staining Method among Hematoxylin and Eosin, Giemsa and Periodic Acid Schiff-Alcian Blue for the Detection of Helicobacter pylori in Gastric Biopsies

Malaysian Journal of Medical Sciences ◽

10.21315/mjms2020.27.5.6 ◽

2020 ◽

Vol 27 (5) ◽

pp. 53-61

Author(s):

Abdullah Saleh Alkhamiss

Keyword(s):

Helicobacter Pylori ◽

Alcian Blue ◽

National Guard ◽

Periodic Acid ◽

Giemsa Stain ◽

Periodic Acid Schiff ◽

Gastric Biopsies ◽

Hematoxylin And Eosin ◽

H Pylori ◽

Sensitivity Specificity

Background: This study was undertaken to evaluate the preferred method (Giemsa or periodic acid Schiff-Alcian blue [PAS-AB] stains) of detecting Helicobacter pylori (H. pylori) in gastric mucosal biopsies in terms of sensitivity, specificity and applicability. To the best of my knowledge, this is the first report comparing Giemsa and PAS-AB staining for the detection of H. pylori in such biopsies. Methods: The formalin-fixed paraffin-embedded blocks of 49 gastric biopsies from different patients were collected from the archive of anatomical pathology at King Abdulaziz Medical City, National Guard, Riyadh, Saudi Arabia. From each block, three slides were prepared and analysed using the hematoxylin and eosin (H&E), Giemsa and PAS-AB stains to detect the presence/absence of H. pylori, and the results were compared in terms of sensitivity, specificity and applicability. Results: The majority of the biopsies in this study showed antrum-type gastric mucosa. Only 15 biopsies showed active gastritis, whereas the rest showed chronic gastritis. Three biopsies showed intestinal metaplasia. All were detected by PAS-AB stain, but only two-thirds were detected by H&E stain. Fifteen gastric biopsies showed H. pylori infection in general and in 13 of them, active gastritis cases were discovered. Fourteen out of these 15 H. pylori infection cases were detected by Giemsa stain, whereas only 13 cases were detected by H&E stain. PAS-AB stain showed the worst results since it demonstrated only 40% sensitivity and 67.65% specificity in H. pylori detection. Conclusion: Giemsa stain has better sensitivity and specificity in gastric H. pylori infection detection than PAS-AB. Therefore, using PAS-AB stain to detect H. pylori infection is not recommended.

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Good diagnostic accuracy of a chemiluminescent immunoassay in stool samples for diagnosis of Helicobacter pylori infection in patients with dyspepsia

Journal of Investigative Medicine ◽

10.1136/jim-2015-000004 ◽

2016 ◽

Vol 64 (2) ◽

pp. 388-391 ◽

Cited By ~ 6

Author(s):

María José Ramírez-Lázaro ◽

Josep Lite ◽

Sergio Lario ◽

Pepa Pérez-Jové ◽

Antònia Montserrat ◽

...

Keyword(s):

Helicobacter Pylori ◽

Gold Standard ◽

Rapid Urease Test ◽

Diagnostic Reliability ◽

Predictive Values ◽

Urease Test ◽

Stool Samples ◽

H Pylori ◽

Good Diagnostic Accuracy ◽

Sensitivity Specificity

Laboratory-based chemiluminescence immunoassays (CLIA) are widely used in clinical laboratories. Some years ago, a CLIA test was developed for the detection of Helicobacter pylori in stool samples, known as LIAISON H. pylori SA, but little information on its use has been reported. To evaluate the accuracy of the LIAISON H. pylori SA assay for diagnosing H. pylori infection prior to eradication treatment. Diagnostic reliability was evaluated in 252 untreated consecutive patients with dyspepsia. The gold standard for diagnosing H. pylori infection was defined as the concordance of the rapid urease test (RUT), histopathology and urea breath test (UBT). The CLIA assay was performed according to the manufacturer's instructions. Sensitivity, specificity, positive and negative predictive values, and 95% CIs were calculated. According to the gold standard selected, 121 patients were positive for H. pylori infection and 131 negative. LIAISON H. pylori SA had a sensitivity of 90.1% and a specificity of 92.4%, with positive and negative predictive values of 91.6% and 90.1%, respectively. The accuracy of the LIAISON H. pylori SA chemiluminescent diagnostic assay seems comparable to that of ELISA or the best-performing LFIAs. Its sensitivity and specificity, however, seem slightly lower than those of histology, RUT or UBT. The advantages of the assay are that it is cheap, automated, and minimally labor-intensive.

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European Registry on Helicobacter pylori Management: single-capsule bismuth quadruple therapy is effective in real-world clinical practice

United European Gastroenterology Journal ◽

10.1177/2050640620972615 ◽

2020 ◽

pp. 205064062097261

Author(s):

Olga P Nyssen ◽

Angeles Perez-Aisa ◽

Manuel Castro-Fernandez ◽

Rinaldo Pellicano ◽

Jose M. Huguet ◽

...

Keyword(s):

Helicobacter Pylori ◽

Clinical Practice ◽

Proton Pump Inhibitor ◽

Proton Pump ◽

Real World ◽

Case Report Form ◽

First Line ◽

Quadruple Therapy ◽

Intention To Treat ◽

H Pylori

Background There has been resurgence in the use of bismuth quadruple therapy (proton pump inhibitor, bismuth, tetracycline and metronidazole) for treating Helicobacter pylori infection thanks to a three-in-one single-capsule formulation. Objective To evaluate the effectiveness and safety of the single-capsule bismuth quadruple therapy. Methods Data were collected in a multicentre, prospective registry of the clinical practice of gastroenterologists on the management of H. pylori infection, where patients were registered at the Asociación Española de Gastroenterología REDCap database on an electronic case report form until January 2020. Effectiveness by modified intention-to-treat and per-protocol as well as multivariable analysis were performed. Independent factors evaluated were: age, gender, indication, compliance, proton pump inhibitor dose and treatment line. Results Finally, 2100 patients were prescribed single-capsule bismuth quadruple therapy following the technical sheet (i.e. three capsules every 6 hours for 10 days). The majority of these patients were naive (64%), with an average age of 50 years, 64% women and 16% with peptic ulcer. An overall modified intention-to-treat effectiveness of 92% was achieved. Eradication was over 90% in first-line treatment (95% modified intention-to-treat, n = 1166), and this was maintained as a rescue therapy, both in second (89% modified intention-to-treat, n = 375) and subsequent lines of therapy (third to sixth line: 92% modified intention-to-treat, n = 236). Compliance was the factor most closely associated with treatment effectiveness. Adverse events were generally mild to moderate, and 3% of patients reported a severe adverse event, leading to discontinuation of treatment in 1.7% of cases. Conclusions Single-capsule bismuth quadruple therapy achieved H. pylori eradication in approximately 90% of patients in real-world clinical practice, both as a first-line and rescue treatment, with good compliance and a favourable safety profile.

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Empirical or susceptibility-guided treatment for Helicobacter pylori infection? A comprehensive review

Therapeutic Advances in Gastroenterology ◽

10.1177/1756284820968736 ◽

2020 ◽

Vol 13 ◽

pp. 175628482096873

Author(s):

Javier P. Gisbert

Keyword(s):

Helicobacter Pylori ◽

Clinical Practice ◽

Helicobacter Pylori Infection ◽

Alternative Methods ◽

First Line ◽

Time Saving ◽

Line Therapy ◽

H Pylori ◽

Guided Therapy

Although susceptibility-guided therapy is frequently recommended for Helicobacter pylori infection, the evidence available to date supporting this strategy is limited. The aim of the present article is to review the advantages and limitations of the susceptibility-guided and the empirical strategies to treat this infection. We performed a bibliographic search to identify studies investigating H. pylori susceptibility-guided therapy. Culture is not the only way to assess antibiotic resistance, as different polymerase chain reaction-based approaches have been developed as alternative methods. For detecting H. pylori antimicrobial resistance, a molecular approach based on a stool sample might enable more convenient, time-saving methods. Unfortunately, the antimicrobial susceptibility cannot be obtained in all cases. Furthermore, antibiotic susceptibility testing in clinical practice yields useful information only for a few antibiotics: clarithromycin, metronidazole, and quinolones. In addition, susceptibility towards clarithromycin and metronidazole in vitro does not necessarily lead to eradication in vivo. In the case of H. pylori therapy failure, we should not re-administer any of the antibiotics against which H. pylori has probably become resistant. Our updated meta-analysis showed that susceptibility-guided treatment is not better than empirical treatment of H. pylori infection in first-line therapy if the most updated quadruple regimens are empirically prescribed, and similar efficacy results were also demonstrated with the two strategies for second-line therapy. Cumulative H. pylori eradication rate with several successive rescue therapies empirically prescribed reaches almost 100%. Finally, the studies that have evaluated the cost-effectiveness of the susceptibility-guided treatment have achieved contradictory results. In summary, we can conclude that the evidence is too limited to support the generalized use of susceptibility-guided therapy for H. pylori treatment in routine clinical practice, either as first-line or as rescue treatment. Nevertheless, it would be recommended that susceptibility tests are performed routinely, even before prescribing first-line treatment, in specialized centers with an interest in H. pylori management.

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