Comparative Efficacy of Various Histological Staining Methods for Demonstration of Helicobacter Pylori in Gastric Biopsies

Comparative Efficacy of Various Histological Staining Methods for Demonstration of Helicobacter Pylori in Gastric Biopsies | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Comparative Efficacy of Various Histological Staining Methods for Demonstration of Helicobacter Pylori in Gastric Biopsies Sujata Thapa, Shreya Shrivastav, Bishal Budha, Bijaya Dhakal, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7142394/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Helicobacter pylori is a spiral, flagellated bacterium having abundant urease production capacity. H. pylori bacterium has been implicated in various gastrointestinal disorders including gastritis, gastroduodenal ulcers, mucosa associated lymphoid tissue lymphoma and gastric cancer. Histology is a gold standard technique for the detection of H. pylori. This study aims to evaluate various histological staining methods used for the demonstration of H. pylori in gastric biopsies. Method This study was conducted in the department of Pathology. A total of 100 gastric biopsies were analyzed for the presence of H. pylori with the use of three different histological stains. The Sensitivity, Specificity, Negative predictive value (NPV), Positive predictive value(PPV) and Diagnostic accuracy (DA) of the stains were calculated. Result H. pylori was detected in total 32 gastric biopsy samples. Males (41.6%) were found to be affected more than females (23%) with middle-aged individuals showing the highest susceptibility. Highest frequency of H. pylori was detected by Giemsa stain followed by Gimenez stain and hematoxylin and eosin (H&E) stain. The result of Gimenez stain was superior to H&E stain and comparable to Giemsa stain. The Sensitivity, Specificity, PPV, NPV and DA of H&E stain was 74.19%, 100%, 100%, 89.61%, 92% and Gimenez stain was 96.7%, 100%, 100%, 98.57% ,99%. Conclusion Giemsa stain has better diagnostic accuracy than traditional H&E stain and Gimenez stain for demonstration of H. pylori in gastric biopsies while Gimenez stain has better Sensitivity, Specificity and diagnostic accuracy than H&E stain. Thus, both Giemsa and Gimenez stains are recommended for the detection of H. pylori in gastric biopsies. Biological sciences/Cancer Health sciences/Diseases Health sciences/Gastroenterology Health sciences/Medical research Biological sciences/Microbiology Helicobacter pylori H and E stain Gimenez Stain Giemsa stain Gastritis Gastric Biopsy Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7142394","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":493053532,"identity":"401da14a-b309-4603-953c-0e0dbf2dad14","order_by":0,"name":"Sujata Thapa","email":"","orcid":"","institution":"Tribhuvan University Teaching Hospital","correspondingAuthor":false,"prefix":"","firstName":"Sujata","middleName":"","lastName":"Thapa","suffix":""},{"id":493053533,"identity":"b5f2107d-c3c3-4fe7-a472-7c1a25eb4d2b","order_by":1,"name":"Shreya Shrivastav","email":"","orcid":"","institution":"Tribhuvan University Teaching Hospital (TUTH)","correspondingAuthor":false,"prefix":"","firstName":"Shreya","middleName":"","lastName":"Shrivastav","suffix":""},{"id":493053534,"identity":"ee4533bc-cde0-4f60-b20d-76eac683b8aa","order_by":2,"name":"Bishal Budha","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA8klEQVRIiWNgGAWjYBACA2YGNgaGggMM/Oz9Dx8ABXj4iNNicIBBsucMswFICxtBLQxQLQY3fNgkQCIEtZizsz978MPgjjzDDd5jlV9z7GTYGJgfPrqBR4tlM4+5YY/BM8PG2X1pt2W3JQMdxmZsnIPPYYd52CR4DA4nMMscMLstuY0ZqIWHTRq/FvZnkn+AWtgkEsyKJbfVE6OFwUwaZAuPRI4Z48dthwlrAfrFTFoG6JcZPMeSpRm3HedhYybgF3P+488k31Tckbc/3nzw489t1fb87M0PH+PTggKYecAkscpBgPEHKapHwSgYBaNgxAAAZEJEqILS20oAAAAASUVORK5CYII=","orcid":"","institution":"Tribhuvan University Teaching Hospital (TUTH)","correspondingAuthor":true,"prefix":"","firstName":"Bishal","middleName":"","lastName":"Budha","suffix":""},{"id":493053535,"identity":"76744e08-c502-4a3c-99d5-c46f25467c3b","order_by":3,"name":"Bijaya Dhakal","email":"","orcid":"","institution":"Tribhuvan University Teaching Hospital","correspondingAuthor":false,"prefix":"","firstName":"Bijaya","middleName":"","lastName":"Dhakal","suffix":""},{"id":493053536,"identity":"9bccf02a-83a8-45a9-bb01-ed2b0fe92d0c","order_by":4,"name":"Sagar Parajuli","email":"","orcid":"","institution":"Tribhuvan University Teaching Hospital","correspondingAuthor":false,"prefix":"","firstName":"Sagar","middleName":"","lastName":"Parajuli","suffix":""},{"id":493053537,"identity":"e713178b-26bb-4575-becb-18795df86ad4","order_by":5,"name":"Sujita Shrestha","email":"","orcid":"","institution":"Tribhuvan University Teaching Hospital","correspondingAuthor":false,"prefix":"","firstName":"Sujita","middleName":"","lastName":"Shrestha","suffix":""}],"badges":[],"createdAt":"2025-07-16 17:23:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7142394/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7142394/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":92926877,"identity":"268a40ed-8c9e-4577-8936-c5f5258d7cae","added_by":"auto","created_at":"2025-10-07 08:09:16","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":530469,"visible":true,"origin":"","legend":"","description":"","filename":"FInalTobesubmittedinBMCinfectious.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7142394/v1_covered_c88ef449-a83b-40d7-b004-c6091a0ca5d2.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Comparative Efficacy of Various Histological Staining Methods for Demonstration of Helicobacter Pylori in Gastric Biopsies","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Helicobacter pylori, H and E stain, Gimenez Stain, Giemsa stain, Gastritis, Gastric Biopsy","lastPublishedDoi":"10.21203/rs.3.rs-7142394/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7142394/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003e\u003cem\u003eHelicobacter pylori\u003c/em\u003e is a spiral, flagellated bacterium having abundant urease production capacity. \u003cem\u003eH. pylori\u003c/em\u003e bacterium has been implicated in various gastrointestinal disorders including gastritis, gastroduodenal ulcers, mucosa associated lymphoid tissue lymphoma and gastric cancer. Histology is a gold standard technique for the detection of \u003cem\u003eH. pylori.\u003c/em\u003e This study aims to evaluate various histological staining methods used for the demonstration of \u003cem\u003eH. pylori\u003c/em\u003e in gastric biopsies.\u003c/p\u003e\u003ch2\u003eMethod\u003c/h2\u003e\u003cp\u003eThis study was conducted in the department of Pathology. A total of 100 gastric biopsies were analyzed for the presence of \u003cem\u003eH. pylori\u003c/em\u003e with the use of three different histological stains. The Sensitivity, Specificity, Negative predictive value (NPV), Positive predictive value(PPV) and Diagnostic accuracy (DA) of the stains were calculated.\u003c/p\u003e\u003ch2\u003eResult\u003c/h2\u003e\u003cp\u003e\u003cem\u003eH. pylori\u003c/em\u003e was detected in total 32 gastric biopsy samples. Males (41.6%) were found to be affected more than females (23%) with middle-aged individuals showing the highest susceptibility. Highest frequency of \u003cem\u003eH. pylori\u003c/em\u003e was detected by Giemsa stain followed by Gimenez stain and hematoxylin and eosin (H\u0026amp;E) stain. The result of Gimenez stain was superior to H\u0026amp;E stain and comparable to Giemsa stain. The Sensitivity, Specificity, PPV, NPV and DA of H\u0026amp;E stain was 74.19%, 100%, 100%, 89.61%, 92% and Gimenez stain was 96.7%, 100%, 100%, 98.57% ,99%.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eGiemsa stain has better diagnostic accuracy than traditional H\u0026amp;E stain and Gimenez stain for demonstration of \u003cem\u003eH. pylori\u003c/em\u003e in gastric biopsies while Gimenez stain has better Sensitivity, Specificity and diagnostic accuracy than H\u0026amp;E stain. Thus, both Giemsa and Gimenez stains are recommended for the detection of \u003cem\u003eH. pylori\u003c/em\u003e in gastric biopsies.\u003c/p\u003e","manuscriptTitle":"Comparative Efficacy of Various Histological Staining Methods for Demonstration of Helicobacter Pylori in Gastric Biopsies","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-31 12:13:46","doi":"10.21203/rs.3.rs-7142394/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c76a1ca6-364d-48a7-a91d-ec178cc0a88d","owner":[],"postedDate":"July 31st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":52353840,"name":"Biological sciences/Cancer"},{"id":52353841,"name":"Health sciences/Diseases"},{"id":52353842,"name":"Health sciences/Gastroenterology"},{"id":52353843,"name":"Health sciences/Medical research"},{"id":52353844,"name":"Biological sciences/Microbiology"}],"tags":[],"updatedAt":"2025-11-06T05:23:31+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-31 12:13:46","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7142394","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7142394","identity":"rs-7142394","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}