Hemagglutination inhibition and nt assays have been utilized for years to investigate influenza immunity, although only a few studies have directly compared the assays influenza antibody detection capabilities, and most of these studies evaluated vaccine. Comparison approaches to measuringinfluenza a virus. Automation of a hemagglutinationinhibition test for parainfluenza 3. Standardization of hemagglutination inhibition assay for in. A wide variety of different viruses possess the ability to agglutinate the erythrocytes of mammalian or avian species. Jun 14, 2012 this video belongs to the section entitled serological tests that is part of the dvd avian influenza sampling procedures and laboratory testing funded by fao and the istituto zooprofilattico.
The ha assay is not an identification assay, as other agents also have hemagglutinating properties. Neither seasonal nor challengebased correlates have been evaluated in. Standardization of the hemagglutination inhibition hai assay for influenza serology is challenging. The test relies on the hemagglutination activity of the influenza ha and the ability of haspecific antibodies to inhibit the virus from agglutinating erythrocytes fig. Influenza antibody titers were detected using hai with 2fold serial dilutions, as described elsewhere,, at the naval medical research center silver spring, md. The standardized hemagglutination ha and hemagglutination inhibition hi. Hemagglutination definition of hemagglutination by the free.
Then select what you want to calculate the inhibition ic50 or induction ic50. Hemagglutination inhibition assays background hi assay example titer is 80. Choose from 19 different sets of hemagglutination flashcards on quizlet. Introduction this supplemental assay method sam describes an in vitro assay method for determining the hemagglutination inhibition hi antibody titer of sera from guinea pigs vaccinated with type a. The standardized hemagglutination test has a reproducibility of 84 to 96% with adenoviruses, rubeola, and the myxoviruses, and 78 to 93%with reoviruses. A simple method for increasing hemagglutinating activity of measles virus tissue culture material is described. This study aimed to assess the prevalence and distribution of hemagglutination inhibition antibodies against antigens of six flavivirus yellow fever virus, ilheus virus, saint louis encephalitis virus, cacipacore virus, bussuquara virus and. The hemagglutination assay is a simple and easy method to obtain semiquantitative data on the sugar binding and specificity of a lectin. One of the techniques used to reach this conclusion is the hemagglutination inhibition hi assay. Ha and the hemagglutination inhibition assay hi or hai were developed in 194142 by american virologist george hirst as methods for quantifying the relative concentration of viruses, bacteria, or antibodies ha and hi apply the process of hemagglutination, in which sialic acid receptors on the surface of red. As each virus molecule binds to multiple rbcs, a latticestructure will form.
Determination of hemagglutinationinhibition titers to. Learn hemagglutination with free interactive flashcards. The classical hemagglutination inhibition chai assay has been in widespread use because of its utility. A viral suspension may agglutinate the red blood cells, thus preventing them from settling out of suspension.
Comparison of the hemagglutination inhibition procedure. Detection of hemagglutinating viruses uncontrolled copy 1. Improvements to the hemagglutination inhibition test for serological assessment of recombinant fowlpoxh5avianinfluenza vaccination in chickens and its use along with an agar gel immunodiffusion test for differentiating infected from noninfected vaccinated animals. The hemagglutination inhibition hai assay is the primary method for determining quantitative antibody titers for influenza virus and is widely used both for licensure of vaccines and for seroepidemiologic studies examining protection in populations 1, 3. After incubation, the degree of agglutination is determined.
So, it does not matter which software you are using, you have. For serological applications, the lack of standardization in. Spotforming cells were measured and counted with automatic elispot reader aid ispot reader and aid software version 5. Hemagglutination inhibition test is widely used for the diagnosis of infection caused by orthomyxoviruses influenza, paramyxoviruses measles, mumps, mononucleosis, abrovirusestogaviruses including rubella, flaviviruses, and bunyaviruses. Some viruses can bind to molecules present on red blood cells, a process called hemagglutination. This will block the receptor sites of the antibodies, thus inhibiting their capacity to cause hemagglutination. Hatchette,f,g,h on behalf of the public health agency of canada and canadian institutes of. Prior to testing, each serum was treated with receptor destroying enzyme cholera filtrate, sigma, germany to inactivate nonspecific inhibitors achieving a final serum dilution of 1. By treatment with tween 80 and ether a 4 to 8fold increase in titer was obtained with virus material grown in a human embryonic cell line lu 106 and an 8 to 32fold increase when virus was grown in dog kidney tc.
Adaptation and limitations of established hemagglutination inhibition assays for the detection of porcine antiswine in. Hemagglutinationinhibition assay for influenza virus. Indevr launches automated hemagglutination analysis system. Manual versus automated readers for hemagglutination. The national center for biomedical ontology was founded as one of the national centers for biomedical computing, supported by the nhgri, the nhlbi, and the nih common fund under grant u54hg004028. The hi titer is defined as the inverse dilution of the last dilution which completely inhibited hemagglutination. Adaptation and limitations of established hemagglutination. The hemagglutination assay or haemagglutination assay ha and the hemagglutination inhibition assay hi or hai were developed in 194142 by american virologist george hirst as methods for quantifying the relative concentration of viruses, bacteria, or antibodies.
Hemagglutination inhibition assay hai this test is based on the inhibition of viral agglutination by a specific antibody and can be used for virus identification or for assay of antibodies. Hemagglutination assay for influenza virus springerlink. Also feel free to ask me any question regarding the protocol you might have like. Hemagglutination inhibition and nt assays have been utilized for years to investigate influenza immunity, although only a few studies have directly compared the assays influenza antibody detection capabilities, and most of these studies evaluated vaccinederived immuni ty. This process can be inhibited by antibodies directed against the virus, blocking the virus from binding to the red blood cells. Hemagglutination inhibition hi assay guide to services. The hemagglutination inhibition hi test is a serological assay that measures the titer of specific antibodies in the sera and is the most common serological assay used to detect antiinfluenza antibodies in swine sera. Both conceptually and operationally, the manual reading step is a potential bottleneck and point of risk for. A hemagglutination inhibition assay 38 was performed to determine hemagglutination inhibition titers. The actual animal species whose erythrocytes could be agglutinated depends on the actual virus. Quizlet flashcards, activities and games help you improve your grades. An optimized hemagglutination inhibition hi assay to.
Find out information about hemagglutination inhibition. Automated imaging and analysis of the hemagglutination inhibition. Automated analysis of hemagglutination and hemagglutination. The hemagglutination assay or haemagglutination assay ha and the hemagglutination inhibition assay hi or hai were developed in 194142 by american virologist george hirst as methods for quantifying the relative concentration of viruses, bacteria, or antibodies ha and hi apply the process of hemagglutination, in which sialic acid receptors on the surface of red blood cells rbcs bind. For influenza virus, hi assay is used to identify the hemagglutinin ha subtype of an unknown isolate or the. Automated interpretation of influenza hemagglutination. Automated imaging and analysis of the hemagglutination inhibition assay. The ha assay can be easily modified to determine the level of antibodies to influenza virus present in serum samples. Naturallyoccurring inhibitors of hemagglutination by influenza viruses. It is best to express an average influenza hi titer as a geometric mean. For additional information regarding classification of specimens for the purposes of shipping, consult either part 2 appendix 3 of the tdg regulations or section 3.
Hemagglutination inhibition definition of hemagglutination. Standardized viral hemagglutination and hemagglutination. Fractional dilution procedure for precise titration of hemagglutinating viruses and hemagglutination inhibiting antibodies. Indevr cypher oneautomated hemagglutination analyzer. Hemagglutination ha, hemagglutination inhibition hi and complement fixation assays are all used for the purpose of quantification of abs, bacteria or viruses using titration. The results of an hai assay are based on an endpoint titration where the titers are generally manually interpreted and recorded by a welltrained expert. Virusspecific antibody secreting cell, memory bcell, and. While preparation can be done on tecan evo platform today, analysis is still done manually by inspecting the resulting microplates and assessing the results by eye. The hi assay is used to measure the level of antibodies in a patients serum that will prevent the agglutination of susceptible erythrocytes by inactivated antigens that are able to attach to the erythrocyte receptors on red blood cells.
Ha is a method of quantification for viruses or bacteria by hemagglutination. Automated imaging and analysis of the hemagglutination. Our new crystalgraphics chart and diagram slides for powerpoint is a collection of over impressively designed datadriven chart and editable diagram s guaranteed to impress any audience. The assay relies on the ability of the hemagglutinin protein on the surface of influenza virus to bind to sialic acids on. In this study, we standardized hai assay materials. It is an easy, simple and rapid method which can be applied. This assay is still used as a standard for measuring the efficacy of influenza vaccines and potency of neutralizing antibodies. Nov 29, 20 the hemagglutination assay or haemagglutination assay. Hemagglutination inhibition article about hemagglutination. The hemagglutination assay or haemagglutination assay ha and the hemagglutination inhibition assay hi or hai were developed in 194142 by american. Current procedural terminology hemagglutination inhibition. Next generation cypher one software increases functionality and ensures data integrity for the standardization of hemagglutination and hemagglutination inhibition.
Animate characters in real time with adobe character animator. Meijer et al 2006 use the ability of antigen, avian influenza virus h7n7 to bind and agglutinate turkey or horse red blood cells to investigate presence of antibodies to h7n7 in patients serum which prevents the hemagglutination of red blood cells by the virus. Hemagglutination inhibition test was conducted by adding 25ul protein to 25ul yeast mannan solution and finally 25ul of blood was added after 1h incubation at room temperature. Research article electron microscopy, hemagglutination inhibition. Thehemagglutination inhibition haitest, whichdetects subtypespecific antibodies against hemagglutinin ha, is commonly used for serologic diagnosis of influenza virus infection andfor determination ofsusceptibility to influenza virusin epidemiologic andvaccinestudies. The hemagglutination assay or haemagglutination assay. The hemagglutination inhibition assay was performed as previously described lenette and schmidt, 1979. For individuals, two tests can have the same result based upon chance alone. Learn vocabulary, terms, and more with flashcards, games, and other study tools. An easy way to perform the hemagglutination inihibition or hi test.
The hemagglutination assay ha is a tool used to screen cell culture isolates or amnioallantoic fluid harvested from embryonated chicken eggs for hemagglutinating agents, such as type a influenza. The relationship between the hemagglutinationinhibition test. Hemagglutination ha assay protocol the hemagglutination assay is a method for titering influenza viruses based on their ability to attach to molecules present on the surface of red blood cells. Hemagglutination inhibition hi assay is a classical laboratory procedure for the classification or subtyping of hemagglutinating viruses.
An active lectin agglutinates erythrocytes by recognizing a carbohydrate on the cell surface and forming a crosslinked network in suspension. Hemagglutination definition of hemagglutination by merriam. Hemagglutination inhibition an overview sciencedirect. Therefore hemagglutination is inhibited when antibodies are. Centers for disease control and prevention have determined that some adults have serum crossreactive antibodies to the new influenza h1n1 virus. Find all of the stateoftheart solutions you need to cost effectively develop and manufacture product. Immunology lab agglutination inhibition and hemagglutination inhibition study guide by smf88 includes 6 questions covering vocabulary, terms and more. Find out information about hemagglutination inhibition test. Hemagglutination definition is agglutination of red blood cells.
T1 determination of hemagglutinationinhibition titers to influenza a virus in porcine sera by use of an enzymelinked immunosorbent assay. How can i calculate ic50 value from percent inhibition. Founded in 1911, it has grown steadily from a small civil servants school to a modern comprehensive university, providing instruction for about 14,000 students in twentyfive facilities. The hemagglutination inhibition assay hai is widely used to evaluate vaccineinduced antibody responses as well as to antigenically characterize influenza viruses. Indevr launches nextgeneration software for the cypher. In hemagglutination or hemagglutination inhibition assay, hia, or passive hemagglutination assay, pha, rbcs generally sheep, which have been coated or coupled with antigen thus called sensitized rbcs, are incubated with antibody and sample. Hemagglutination, or haemagglutination, is a specific form of agglutination that involves red blood cells rbcs. Hemagglutination inhibition hi has been a reliable method for determining porcine antibody. Synonyms for hemagglutination inhibition in free thesaurus. Traditionally, serum has been the specimen of choice for hai assays, but a desire to reduce the amount of blood collected during studies and the availability of plasma in archived.
Agglutination inhibition test pregnancy test youtube. Poor reproducibility of hai results from one laboratory to another is widely cited, limiting comparisons between candidate vaccines in different clinical trials and posing challenges for licensing authorities. Hemagglutination inhibition synonyms, hemagglutination. Aug 27, 2016 hemagglutination inhibition and nt assays have been utilized for years to investigate influenza immunity, although only a few studies have directly compared the assays influenza antibody detection capabilities, and most of these studies evaluated vaccine.
Animation software, animation tools and apps adobe. Kappa is a measure of agreement between two tests that corrects for this chance agreement. The first known use of hemagglutination was in 1907. An optimized hemagglutination inhibition hi assay to quantify influenzaspecific antibody titers lukas kaufmann 1, mohammedyaseen syedbasha 1, dominik vogt 1, yvonne hollenstein 1, julia hartmann 1, janina e.
I got a list of influenzaspecific antibody titers from a hemagglutination inhibition assay. Hemagglutination inhibition tests are simple, sensitive, inexpensive, and rapid and therefore are often the method of choice for assaying antibodies to influenza a virus. Elisa was performed to measure binding antibodies to fulllength hemagglutinin, hemagglutinin. This hemagglutination may be inhibited if isolated antigens of the erythrocyte membrane are first incubated with the antiserum. Supplemental assay method for conducting the hemagglutination. Chart and diagram slides for powerpoint beautifully designed chart and diagram s for powerpoint with visually stunning graphics and animation effects. May 27, 2016 hemagglutination inhibition test was conducted by adding 25ul protein to 25ul yeast mannan solution and finally 25ul of blood was added after 1h incubation at room temperature. Hemagglutinin inhibition assay with swine sera springerlink. Indevr has validated cypherone using both turkey and guinea pig rbcs in u bottom and v bottom plates for both ha and hai. The inhibition is specific and is widely used for virus identification and for antibody determination. Comparative analysis of hemagglutination inhibition titers.
Introduction this testing protocol pro describes the procedures used to detect and quantitate the presence of extraneous newcastle disease virus ndv or other hemagglutinating ha viruses that may be present in veterinary vaccines or master seeds ms other than live ndv ms or vaccines. Comparison of hemagglutination inhibition assay, an elisa. This category has the following 3 subcategories, out of 3 total. Definition, synonyms, translations of hemagglutination inhibition hi test by the free dictionary. An automated hemagglutinationinhibition hi test for the shipping fever strain sf4 of parainfluenza 3 antibody in bovine sera was developed and compared. An enzymelinked immunosorbent assay elisa and the hemagglutination inhibition hi test were used to evaluate the response of laboratory rats to experimental infection with pneumonia virus of mice. This video belongs to the section entitled serological tests that is part of the dvd avian influenza sampling procedures and laboratory testing funded by fao and the istituto zooprofilattico. The presence of virus in infected cell cultures can be detected by hemagglutination. Food and drug administration fda of automated interpretation of influenza hemagglutination inhibition hai assays.
Seroprevalence of flaviviruses antibodies in water. Hemagglutination inhibition antibody titers as a correlate. Rapid automated analysis of hemagglutination and hemagglutination inhibition assays to standardize analysis and provide a digital, traceable record. Test and partial sequencing analysis of influenza a virus samples isolated from wild birds in sao paulo state, brazil. Shipping of specimens shall be done by a tdg certified individual in accordance with tdg regulations.
Influenzaspecific hemaggluitination inhibition hai antibody titer, an indicator of immunity to influenza, is often used to measure exposure to influenza in surveillance and immunogenicity studies. Automated analysis of hemagglutination and hemagglutination inhibition assays. Standardization of hemagglutination inhibition assay for. The allantoic fluid from each surviving egg was tested for the presence of hemagglutinating agent by using ha test and was further confirmed through virus neutralization hemagglutination inhibition test.
The serum samples were evaluated by hemagglutination inhibition hi assay, as previously described. William james explains the principles of the haemagglutination inhibitation assa. The elisa procedure was more sensitive than the hi test and detected very low levels of antibodies early in the course of infection. William james explains how the haemagglutination assay can be used to measure the influenza a virus titre in a sample.
1579 1443 460 113 1219 1119 1087 503 1259 1323 1598 1470 690 1083 105 27 850 671 1349 828 1398 101 1025 1100 25 1283 1059 969 1064 526 425 205 526 441 597 1382 1199 788 924 719 574