{"id":799,"date":"2022-07-04T20:45:32","date_gmt":"2022-07-04T15:00:32","guid":{"rendered":"https:\/\/immunologynotes.com\/?p=397"},"modified":"2022-09-05T13:51:47","modified_gmt":"2022-09-05T08:06:47","slug":"radial-immunodiffusion","status":"publish","type":"post","link":"https:\/\/microbenotes.com\/radial-immunodiffusion\/","title":{"rendered":"Radial Immunodiffusion- Principle, Procedure, Results, Uses"},"content":{"rendered":"<p><span style=\"color: #000000;\"><strong>Immuno-diffusion<\/strong> is a technique for the detection or measurement of antibodies and antigens by their precipitation which involves diffusion through a substance such as agar or gel agarose. Simply, it denotes precipitation in gel.<\/span><\/p>\n<p><span style=\"color: #000000;\">It refers to any of the several techniques for obtaining a precipitate between an antibody and its specific antigen.<\/span><\/p>\n<p><span style=\"color: #000000;\">This can be achieved by:<\/span><\/p>\n<ol>\n<li><span style=\"color: #000000;\">a) Suspending antigen\/antibody in a gel and letting the other migrate through it from a well or,<\/span><\/li>\n<li><span style=\"color: #000000;\">b) Letting both antibody and antigen migrate through the gel from separate wells such that they form an area of precipitation.<\/span><\/li>\n<\/ol>\n<p><span style=\"color: #000000;\">Based on the method employed, immuno-diffusion may be:<\/span><\/p>\n<ol>\n<li><span style=\"color: #000000;\"><strong>Radial immunodiffusion<\/strong><\/span><\/li>\n<li><span style=\"color: #000000;\"><strong> Ouchterlony Double Diffusion<\/strong><\/span><\/li>\n<\/ol>\n<p><span style=\"color: #000000;\"><strong>Radial immunodiffusion\u00a0(RID)<\/strong> or <strong>Mancini method<\/strong> is also known as <strong>Mancini immunodiffusion<\/strong> or <strong>single radial immunodiffusion assay<\/strong>. It is a single diffusion technique whereby a\u00a0solution\u00a0containing\u00a0the\u00a0antigen\u00a0is\u00a0placed into wells in\u00a0a\u00a0gel or agar surface evenly\u00a0impregnated\u00a0with\u00a0antibody. The\u00a0diameter\u00a0of\u00a0the\u00a0ring\u00a0that\u00a0precipitates\u00a0around\u00a0the well as a result of antigen antibody reaction\u00a0corresponds to the amount of\u00a0antigen\u00a0in\u00a0the\u00a0solution.<\/span><\/p>\n<div id=\"ez-toc-container\" class=\"ez-toc-v2_0_65 counter-hierarchy ez-toc-counter ez-toc-custom ez-toc-container-direction\">\n<div class=\"ez-toc-title-container\">\n<p class=\"ez-toc-title \" >Table of Contents<\/p>\n<span class=\"ez-toc-title-toggle\"><a href=\"#\" class=\"ez-toc-pull-right ez-toc-btn ez-toc-btn-xs ez-toc-btn-default ez-toc-toggle\" aria-label=\"Toggle Table of Content\"><span class=\"ez-toc-js-icon-con\"><span class=\"\"><span class=\"eztoc-hide\" style=\"display:none;\">Toggle<\/span><span class=\"ez-toc-icon-toggle-span\"><svg style=\"fill: #0a0a0a;color:#0a0a0a\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" class=\"list-377408\" width=\"20px\" height=\"20px\" viewBox=\"0 0 24 24\" fill=\"none\"><path d=\"M6 6H4v2h2V6zm14 0H8v2h12V6zM4 11h2v2H4v-2zm16 0H8v2h12v-2zM4 16h2v2H4v-2zm16 0H8v2h12v-2z\" fill=\"currentColor\"><\/path><\/svg><svg style=\"fill: #0a0a0a;color:#0a0a0a\" class=\"arrow-unsorted-368013\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"10px\" height=\"10px\" viewBox=\"0 0 24 24\" version=\"1.2\" baseProfile=\"tiny\"><path d=\"M18.2 9.3l-6.2-6.3-6.2 6.3c-.2.2-.3.4-.3.7s.1.5.3.7c.2.2.4.3.7.3h11c.3 0 .5-.1.7-.3.2-.2.3-.5.3-.7s-.1-.5-.3-.7zM5.8 14.7l6.2 6.3 6.2-6.3c.2-.2.3-.5.3-.7s-.1-.5-.3-.7c-.2-.2-.4-.3-.7-.3h-11c-.3 0-.5.1-.7.3-.2.2-.3.5-.3.7s.1.5.3.7z\"\/><\/svg><\/span><\/span><\/span><\/a><\/span><\/div>\n<nav><ul class='ez-toc-list ez-toc-list-level-1 ' ><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-1\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/#objectives-of-radial-immunodiffusion\" title=\"Objectives of Radial Immunodiffusion\">Objectives of Radial Immunodiffusion<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-2\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/#principle-of-radial-immunodiffusion\" title=\"Principle\u00a0of Radial Immunodiffusion\">Principle\u00a0of Radial Immunodiffusion<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-3\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/#procedure-of-radial-immunodiffusion\" title=\"Procedure\u00a0of Radial Immunodiffusion\">Procedure\u00a0of Radial Immunodiffusion<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-4\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/#result-interpretation-of-radial-immunodiffusion\" title=\"Result Interpretation\u00a0of Radial Immunodiffusion\">Result Interpretation\u00a0of Radial Immunodiffusion<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-5\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/#-applications-of-radial-immunodiffusion\" title=\"\u00a0Applications\u00a0of Radial Immunodiffusion\">\u00a0Applications\u00a0of Radial Immunodiffusion<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-6\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/#advantages-of-radial-immunodiffusion\" title=\"Advantages\u00a0of Radial Immunodiffusion\">Advantages\u00a0of Radial Immunodiffusion<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-7\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/#limitations-of-radial-immunodiffusion\" title=\"Limitations\u00a0of Radial Immunodiffusion\">Limitations\u00a0of Radial Immunodiffusion<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-8\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/#references\" title=\"References\">References<\/a><\/li><\/ul><\/nav><\/div>\n<h2><span class=\"ez-toc-section\" id=\"objectives-of-radial-immunodiffusion\"><\/span><span style=\"color: #000000;\"><strong>Objectives of Radial Immunodiffusion<\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><span style=\"color: #000000;\">The Mancini immunodiffusion test may be carried out with one or more of the following objectives:<\/span><\/p>\n<ol>\n<li><span style=\"color: #000000;\">To detect antigen-antibody complexes.<\/span><\/li>\n<li><span style=\"color: #000000;\">Describe the circumstances under which antigen-antibody complexes precipitate out.<\/span><\/li>\n<li><span style=\"color: #000000;\">Determine relative concentration of antigens.<\/span><\/li>\n<\/ol>\n<h2><span class=\"ez-toc-section\" id=\"principle-of-radial-immunodiffusion\"><\/span><span style=\"color: #000000;\"><strong>Principle\u00a0of Radial Immunodiffusion<\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><span style=\"color: #000000;\">Radial immuno-diffusion is a type of precipitation reaction. It is thus based on the principles of the\u00a0precipitin\u00a0curve which states that antigen-antibody interact forming visible cross-linked precipitate when the proper ratio of antigen to antibody is present. In the test, antibody is incorporated into agar and poured into a glass plate to form a uniform layer. Circular wells are cut into the agar and antigen is introduced into the wells. Specific antigens to the impregnated antibodies diffuse through the agar in all directions from the well and react with the antibody present forming visible precipitate or a precipitin ring. Ring shaped bands of precipitates from concentrically around the well indicating reaction. The\u00a0diameter\u00a0of\u00a0the\u00a0precipitate ring formed, corresponds to the amount of\u00a0antigen\u00a0in\u00a0the\u00a0solution.<\/span><\/p>\n<h2><span class=\"ez-toc-section\" id=\"procedure-of-radial-immunodiffusion\"><\/span><span style=\"color: #000000;\"><strong>Procedure\u00a0of Radial Immunodiffusion<\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<ol>\n<li><span style=\"color: #000000;\">An agar containing an appropriate antiserum (antibody) is poured in plates.<\/span><\/li>\n<li><span style=\"color: #000000;\">Carefully circular wells are cut and removed from the plates.<\/span><\/li>\n<li><span style=\"color: #000000;\">A single or series of standards containing known concentration of antigen are placed in separate wells, while control and \u201cunknown\u201d samples are placed in other remaining wells.<\/span><\/li>\n<li><span style=\"color: #000000;\">As the antigen diffuses radially, a ring of precipitate will form in the area of optimal antigen &#8211; antibody concentration.<\/span><\/li>\n<li><span style=\"color: #000000;\">The ring diameters are measured and noted.<\/span><\/li>\n<li><span style=\"color: #000000;\">A standard curve is prepared using the ring diameters of the standards versus their concentrations. This curve is then used to determine the concentration of the control and unknown samples.<\/span><\/li>\n<\/ol>\n<h2><span class=\"ez-toc-section\" id=\"result-interpretation-of-radial-immunodiffusion\"><\/span><span style=\"color: #000000;\"><strong>Result Interpretation\u00a0of Radial Immunodiffusion<\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"aligncenter size-full wp-image-398\" src=\"https:\/\/microbenotes.com\/wp-content\/uploads\/2018\/07\/Result-Interpretation-of-Radial-Immunodiffusion.jpg\" alt=\"Result Interpretation\u00a0of Radial Immunodiffusion\" width=\"800\" height=\"420\" \/><\/p>\n<ol>\n<li><span style=\"color: #000000;\">The presence of a precipitin ring around the antigen wells indicate specific antigen-antibody interaction.<\/span><\/li>\n<li><span style=\"color: #000000;\">Absence of precipitin ring suggest absence of reaction.<\/span><\/li>\n<li><span style=\"color: #000000;\">The greater the amount of antigen in the well, the farther the ring will form from the well.<\/span><\/li>\n<\/ol>\n<h2><span class=\"ez-toc-section\" id=\"-applications-of-radial-immunodiffusion\"><\/span><span style=\"color: #000000;\"><strong>\u00a0Applications\u00a0of Radial Immunodiffusion<\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<ul>\n<li><span style=\"color: #000000;\">Immuno-diffusion\u00a0techniques are mostly used in\u00a0immunology\u00a0to determine the quantity or concentration of an\u00a0antigen\u00a0in a sample.<\/span><\/li>\n<li><span style=\"color: #000000;\">Estimation of the immunoglobulin classes in sera.<\/span><\/li>\n<li><span style=\"color: #000000;\">Estimation of IgG, IgM antibodies in sera to influenza viruses.<\/span><\/li>\n<\/ul>\n<p><span style=\"color: #000000;\"><strong>Other applications include:<\/strong><\/span><\/p>\n<ul>\n<li><span style=\"color: #000000;\">To determine relative concentrations of antibodies in serum.<\/span><\/li>\n<li><span style=\"color: #000000;\">Estimate serum transferrin and alpha-feroprotein.<\/span><\/li>\n<li><span style=\"color: #000000;\">To compare properties of two different antigens.<\/span><\/li>\n<li><span style=\"color: #000000;\">To determine the relative purity of an antigen preparation<\/span><\/li>\n<li><span style=\"color: #000000;\">For disease diagnosis<\/span><\/li>\n<li><span style=\"color: #000000;\">Serological surveys<\/span><\/li>\n<\/ul>\n<h2><span class=\"ez-toc-section\" id=\"advantages-of-radial-immunodiffusion\"><\/span><span style=\"color: #000000;\"><strong>Advantages\u00a0of Radial Immunodiffusion<\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<ol>\n<li><span style=\"color: #000000;\">Precipitation in gels is believed to provide more specific and sensitive results than other methods available.<\/span><\/li>\n<li><span style=\"color: #000000;\">The reaction is in the form of bands of precipitation and can be stained for better viewing as well as preservation.<\/span><\/li>\n<li><span style=\"color: #000000;\">If a large number of antigens are present, each antigen-antibody reaction will give rise to a separate line of precipitation.<\/span><\/li>\n<li><span style=\"color: #000000;\">This technique also indicates identity, cross reaction and non identity between different antigens.<\/span><\/li>\n<\/ol>\n<h2><span class=\"ez-toc-section\" id=\"limitations-of-radial-immunodiffusion\"><\/span><span style=\"color: #000000;\"><strong>Limitations\u00a0of Radial Immunodiffusion<\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<ol>\n<li><span style=\"color: #000000;\">Long reaction time (18-48 hours)<\/span><\/li>\n<li><span style=\"color: #000000;\">It has also been proposed that the results of Mancini&#8217;s test is influenced by the presence bound metal cations in the test samples (protein).<\/span><\/li>\n<li><span style=\"color: #000000;\">Single diffusion menthod of precipitation is considered relatively wasteful than other methods.<\/span><\/li>\n<li><span style=\"color: #000000;\">The test has been recently replaced by more sensitive and automated methods, such as nephelometry and enzyme-linked immunosorbent assays.<\/span><\/li>\n<\/ol>\n<h2><span class=\"ez-toc-section\" id=\"references\"><\/span><span style=\"color: #000000;\"><strong>References<\/strong><\/span><span class=\"ez-toc-section-end\"><\/span><\/h2>\n<ol>\n<li><span style=\"color: #000000;\">Stephan E. Ritzmann; Radial Immunodiffusion Revisited. Part 1,\u00a0Laboratory Medicine, Volume 9, Issue 7, 1 July 1978, Pages 23\u201334.<\/span><\/li>\n<li><span style=\"color: #000000;\">https:\/\/www.slideshare.net\/prashanthkumarguddeti\/antigen-antibody-interactions-63139960<\/span><\/li>\n<li><span style=\"color: #000000;\">Lydyard, P.M., Whelan,A.,&amp; Fanger,M.W. (2005).Immunology (2 ed.).London: BIOS Scientific Publishers.<\/span><\/li>\n<li><span style=\"color: #000000;\">Parija S.C. (2012). Textbook of Microbiology &amp; Immunology.(2 ed.). India: Elsevier India.<\/span><\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"<p>Immuno-diffusion is a technique for the detection or measurement of antibodies and antigens by their precipitation which involves diffusion through a substance such as agar or gel agarose. Simply, it &#8230; <a title=\"Radial Immunodiffusion- Principle, Procedure, Results, Uses\" class=\"read-more\" href=\"https:\/\/microbenotes.com\/radial-immunodiffusion\/\" aria-label=\"Read more about Radial Immunodiffusion- Principle, Procedure, Results, Uses\">Read more<\/a><\/p>\n","protected":false},"author":1,"featured_media":719,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1026],"tags":[1235,1316,1317,1393,1394,1395,1396,1397,1398,1399,1400,1418,1454],"custom":{"td_video":"","featured_image":"https:\/\/microbenotes.com\/wp-content\/uploads\/2018\/07\/Result-Interpretation-of-Radial-Immunodiffusion.jpg","author":{"name":"Sagar Aryal","avatar":"https:\/\/secure.gravatar.com\/avatar\/862472ea06500d4318f1ad82addb608e?s=96&d=mm&r=g"},"categories":[{"term_id":1026,"name":"Immunology","slug":"immunology","term_group":0,"term_taxonomy_id":1026,"taxonomy":"category","description":"<strong>Immunology is an emerging branch of medical science that deals with studies related to different aspects of the immune system like the cells, structure, function, response against antigens, and disorders.<\/strong>\r\n<ul>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">The immune system consists of a collection of cells, chemicals, processes, and mechanisms that function to protect the body from foreign antigens, such as microbes (organisms such as bacteria, fungi, and parasites), viruses, cancer cells, and toxins.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Beyond, the structural and chemical barriers which protect us from infection, the immune system consists of two lines of defense; innate immunity and adaptive immunity.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">The innate immunity represents the first line of defense against the antigen, and it is an antigen-independent defense mechanism used by the host immediately or within hours of encountering an antigen.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">On the other hand, adaptive immunity is antigen-dependent and antigen-specific and, therefore, includes a lag time between exposure to the antigen and maximal response.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Studies conducted in immunology are related to the measurement of physiological functioning of the immune system in both healthy and disease conditions, malfunctioning of the system in the case of disorders, and physical, chemical, and physiological characteristics of the cells of the immune system.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Immunology is fast becoming an important branch of clinical medicine as it has a close relationship with organ transplantation, oncology, virology, bacteriology, and even dermatology.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Immunology is focused on certain organs of the body like the bone marrow and the lymphatic system and the white blood cells found in the blood.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">These cells or organs producing these cells are responsible, directly or indirectly, for the defense mechanism of the body against a pathogenic agent or other antigens.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">These cells keep on circulating throughout the body via blood or lymph so that they can detect antigens entering the body from different sources.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">An important aspect of immunology is immunotherapy, where components of the immune system or antigens are used to treat a disease or disorder as a form of treatment.<\/span><\/li>\r\n<\/ul>","parent":0,"count":115,"filter":"raw","cat_ID":1026,"category_count":115,"category_description":"<strong>Immunology is an emerging branch of medical science that deals with studies related to different aspects of the immune system like the cells, structure, function, response against antigens, and disorders.<\/strong>\r\n<ul>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">The immune system consists of a collection of cells, chemicals, processes, and mechanisms that function to protect the body from foreign antigens, such as microbes (organisms such as bacteria, fungi, and parasites), viruses, cancer cells, and toxins.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Beyond, the structural and chemical barriers which protect us from infection, the immune system consists of two lines of defense; innate immunity and adaptive immunity.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">The innate immunity represents the first line of defense against the antigen, and it is an antigen-independent defense mechanism used by the host immediately or within hours of encountering an antigen.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">On the other hand, adaptive immunity is antigen-dependent and antigen-specific and, therefore, includes a lag time between exposure to the antigen and maximal response.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Studies conducted in immunology are related to the measurement of physiological functioning of the immune system in both healthy and disease conditions, malfunctioning of the system in the case of disorders, and physical, chemical, and physiological characteristics of the cells of the immune system.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Immunology is fast becoming an important branch of clinical medicine as it has a close relationship with organ transplantation, oncology, virology, bacteriology, and even dermatology.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Immunology is focused on certain organs of the body like the bone marrow and the lymphatic system and the white blood cells found in the blood.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">These cells or organs producing these cells are responsible, directly or indirectly, for the defense mechanism of the body against a pathogenic agent or other antigens.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">These cells keep on circulating throughout the body via blood or lymph so that they can detect antigens entering the body from different sources.<\/span><\/li>\r\n \t<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">An important aspect of immunology is immunotherapy, where components of the immune system or antigens are used to treat a disease or disorder as a form of treatment.<\/span><\/li>\r\n<\/ul>","cat_name":"Immunology","category_nicename":"immunology","category_parent":0}]},"_links":{"self":[{"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/posts\/799"}],"collection":[{"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/comments?post=799"}],"version-history":[{"count":0,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/posts\/799\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/media\/719"}],"wp:attachment":[{"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/media?parent=799"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/categories?post=799"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/microbenotes.com\/wp-json\/wp\/v2\/tags?post=799"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}