Mohammad Javad Rezaei, Maryam Eidi, Seyed Ali Mirhosseini, Rouhollah Kazemi, Mohammad Javad Motamedi, Soghra Khani, Jafar Amani
{"title":"肠出血性大肠杆菌elisa诊断系统的设计。","authors":"Mohammad Javad Rezaei, Maryam Eidi, Seyed Ali Mirhosseini, Rouhollah Kazemi, Mohammad Javad Motamedi, Soghra Khani, Jafar Amani","doi":"10.18502/ijm.v17i2.18388","DOIUrl":null,"url":null,"abstract":"<p><strong>Background and objectives: </strong><i>Escherichia coli (E. coli)</i> O157:H7 is an intestinal pathogen of humans and animals, which causes serious gastrointestinal, urinary tract infection and hemolytic uremic syndrome. Connecting to the host cell is important in pathogenesis. EspA, Intimin and Tir proteins (EIT) are the most important bacterial features in the process of binding. These antigens can be very useful in detecting these bacteria. The aim of this study was to produce recombinant EspA, Intimin and Tir proteins (rEIT) to detect pathogenic <i>E. coli</i> O157:H7 by means of ELISA method.</p><p><strong>Materials and methods: </strong>The <i>eit</i> recombinant gene was expressed using IPTG in <i>E. coli</i> BL21 (DE3) and evaluated by western blotting. The purified rEIT protein was injected to rabbits and mice subcutaneously. Purified antibody was evaluated using indirect, competitive and sandwich ELISA confirming the precise detection of <i>E. coli</i> O157: H7.</p><p><strong>Results: </strong>Indirect, competitive and sandwich ELISA specifically detected <i>E. coli</i> O157:H7 and each methods had the ability to identify more than 10<sup>4</sup>, 10<sup>4</sup>, 10<sup>3</sup> bacteria. The specificity of this method was evaluated by Entroheamoragic <i>E. coli</i>, enterotoxygenic <i>E. coli, Klebsiella pneumoniae, Vibrio cholera</i> and <i>Acinetobacter</i>.</p><p><strong>Conclusion: </strong>These methods are the fastest, most accurate and cost effective methods for diagnosis of <i>E. coli</i> O157: H7, comparing to the conventional methods.</p>","PeriodicalId":14633,"journal":{"name":"Iranian Journal of Microbiology","volume":"17 2","pages":"278-286"},"PeriodicalIF":1.3000,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12053411/pdf/","citationCount":"0","resultStr":"{\"title\":\"Design of ELISA-based diagnostic system for detection of enterohaemorrhagic <i>Escherichia coli</i>.\",\"authors\":\"Mohammad Javad Rezaei, Maryam Eidi, Seyed Ali Mirhosseini, Rouhollah Kazemi, Mohammad Javad Motamedi, Soghra Khani, Jafar Amani\",\"doi\":\"10.18502/ijm.v17i2.18388\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background and objectives: </strong><i>Escherichia coli (E. coli)</i> O157:H7 is an intestinal pathogen of humans and animals, which causes serious gastrointestinal, urinary tract infection and hemolytic uremic syndrome. Connecting to the host cell is important in pathogenesis. EspA, Intimin and Tir proteins (EIT) are the most important bacterial features in the process of binding. These antigens can be very useful in detecting these bacteria. The aim of this study was to produce recombinant EspA, Intimin and Tir proteins (rEIT) to detect pathogenic <i>E. coli</i> O157:H7 by means of ELISA method.</p><p><strong>Materials and methods: </strong>The <i>eit</i> recombinant gene was expressed using IPTG in <i>E. coli</i> BL21 (DE3) and evaluated by western blotting. The purified rEIT protein was injected to rabbits and mice subcutaneously. Purified antibody was evaluated using indirect, competitive and sandwich ELISA confirming the precise detection of <i>E. coli</i> O157: H7.</p><p><strong>Results: </strong>Indirect, competitive and sandwich ELISA specifically detected <i>E. coli</i> O157:H7 and each methods had the ability to identify more than 10<sup>4</sup>, 10<sup>4</sup>, 10<sup>3</sup> bacteria. The specificity of this method was evaluated by Entroheamoragic <i>E. coli</i>, enterotoxygenic <i>E. coli, Klebsiella pneumoniae, Vibrio cholera</i> and <i>Acinetobacter</i>.</p><p><strong>Conclusion: </strong>These methods are the fastest, most accurate and cost effective methods for diagnosis of <i>E. coli</i> O157: H7, comparing to the conventional methods.</p>\",\"PeriodicalId\":14633,\"journal\":{\"name\":\"Iranian Journal of Microbiology\",\"volume\":\"17 2\",\"pages\":\"278-286\"},\"PeriodicalIF\":1.3000,\"publicationDate\":\"2025-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12053411/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Iranian Journal of Microbiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.18502/ijm.v17i2.18388\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Iranian Journal of Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.18502/ijm.v17i2.18388","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Design of ELISA-based diagnostic system for detection of enterohaemorrhagic Escherichia coli.
Background and objectives: Escherichia coli (E. coli) O157:H7 is an intestinal pathogen of humans and animals, which causes serious gastrointestinal, urinary tract infection and hemolytic uremic syndrome. Connecting to the host cell is important in pathogenesis. EspA, Intimin and Tir proteins (EIT) are the most important bacterial features in the process of binding. These antigens can be very useful in detecting these bacteria. The aim of this study was to produce recombinant EspA, Intimin and Tir proteins (rEIT) to detect pathogenic E. coli O157:H7 by means of ELISA method.
Materials and methods: The eit recombinant gene was expressed using IPTG in E. coli BL21 (DE3) and evaluated by western blotting. The purified rEIT protein was injected to rabbits and mice subcutaneously. Purified antibody was evaluated using indirect, competitive and sandwich ELISA confirming the precise detection of E. coli O157: H7.
Results: Indirect, competitive and sandwich ELISA specifically detected E. coli O157:H7 and each methods had the ability to identify more than 104, 104, 103 bacteria. The specificity of this method was evaluated by Entroheamoragic E. coli, enterotoxygenic E. coli, Klebsiella pneumoniae, Vibrio cholera and Acinetobacter.
Conclusion: These methods are the fastest, most accurate and cost effective methods for diagnosis of E. coli O157: H7, comparing to the conventional methods.
期刊介绍：
The Iranian Journal of Microbiology (IJM) is an international, multi-disciplinary, peer-reviewed journal that provides rapid publication of the most advanced scientific research in the areas of basic and applied research on bacteria and other micro-organisms, including bacteria, viruses, yeasts, fungi, microalgae, and protozoa concerning the development of tools for diagnosis and disease control, epidemiology, antimicrobial agents, clinical microbiology, immunology, Genetics, Genomics and Molecular Biology. Contributions may be in the form of original research papers, review articles, short communications, case reports, technical reports, and letters to the Editor. Research findings must be novel and the original data must be available for review by the Editors, if necessary. Studies that are preliminary, of weak originality or merely descriptive as well as negative results are not appropriate for the journal. Papers considered for publication must be unpublished work (except in an abstract form) that is not under consideration for publication anywhere else, and all co-authors should have agreed to the submission. Manuscripts should be written in English.
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