People who have had serious allergic reactions to blood transfusions in the past may be transfused with blood that has been "washed" to remove plasma. The prevalence of interference is generally low in assays containing blocking agents that neutralise or inhibit the interference but is often higher in new, untested immunoassays. Multiple spuriously abnormal thyroid function indices due to heterophilic antibodies. Cross-reactivity, in a general sense, is the reactivity of an observed agent which initiates reactions outside the main reaction expected. [1]:236, Prior to receiving a blood transfusion, individuals are screened for the presence of antibodies against antigens of non-ABO blood group systems. 20. Physicians should be encouraged to communicate specifically with the laboratory about discordance between results and clinical findings. Autoantibodies have been described and can cause interference in both non-immunoassay and immunoassay methods for a number of analytes including macro-enzymes (creatine kinase, amylase), thyroid hormones in both free and total forms,40,52,53 thyroglobulin,52 insulin,54,55 prolactin,56 and testosterone.57 False-positive or false-negative values may arise, depending on whether the autoantibody-analyte complex partitions into the free or the bound analyte fraction. . In this case, the recipient's blood type is simply compared against that of the donor blood, without any need for serologic testing. Certain drugs or other chemicals can give a false positive for another category of drug.[1]. Interference affects a wide range of immunoassay analytes including hormones, tumour markers, drugs, cardiac troponin, and microbial serology. Race published a description of the antiglobulin test (also known as the Coombs test). False increase of cardiac troponin I with heterophilic antibodies. have similar chemical structures. For example, if a person has been transfused with large volumes of donor blood, the results of serologic testing will reflect the antigens on the donor cells and not the person's actual blood type. The anti-human globulin could bind to antibodies already attached to red blood cells and cause them to agglutinate. The patient's plasma agglutinates type A1 and B red blood cells. Cross-reactions are common in blood tests for fluke infections because the different species are closely related. Autoantibodies to thyroid hormones have been reported in patients with Hashimotos thyroiditis, Graves disease, hyperthyroidism after treatment, carcinoma, goitre and non-thyroid autoimmune conditions.52 The prevalence of thyroid hormone autoantibodies is dependent on the detection system and may be as high as 10% in patients with autoimmune disease although only a minority of samples cause interference.52,53 Their presence should be suspected when FT4 and thyroid-stimulating hormone (TSH) results appear to be discordant to the laboratory or the clinician.58 Sakata et al. Analysis of hCG: clinical applications and assay requirements. Cross-reaction is a problem in diagnostic immunoassays where endogenous molecules . [16], In this case, the antibody panel shows that anti-Fya antibodies are present. Binding of cortisol can be minimised by denaturation of the binding protein or by the addition of blocking agent. fulminans can form fucose and an oligosaccharide having considerable cross-reactivity with anti-Dip. the contents by NLM or the National Institutes of Health. Price C, Newman D. In: Price CP, Newman DJ, editors. True anaphylaxis (severe drop in blood pressure, loss of consciousness, resuscitation, acute and severe gastrointestinal symptoms (i.e., >5 times diarrhea), other emergency medical measures) . Kricka LJ. If routine serologic testing for RhD results in a score of 2+ or less, the antiglobulin test can be used to demonstrate the presence of RhD. 445480. Zweig MH, Csako G, Spero M. Escape from blockade of interfering heterophile antibodies in a two-site immunoradiometric assay for thyrotropin. Desprs N, Grant AM. [letter]. Glendenning P, Musk AA, Taranto M, Vasikaran SD. . Weakened expression of ABO antigens may also occur in leukemia and Hodgkin's lymphoma. The Red Cross says a slow in blood donations over the last few weeks is concerning, and they're offering perks for donors. A review. Misleading results from immunoassays of serum free thyroxine in the presence of rheumatoid factor. Acad. [1]:2623, Blood typing can be performed using test tubes, microplates, or blood typing slides. When a pregnant woman has a blood group antibody that can cause HDN, the fetus can be typed for the relevant antigen to determine if it is at risk of developing the disease. Ellis MJ, Livesey JH, Evans MJ. Cold agglutinins can usually be deactivated by warming the sample to 37C (99F) and washing the red blood cells with saline. Miller JJ, Valdes Jr R. Approaches to minimizing interference by cross-reacting molecules in immunoassays. For example, in central hypothyroidism immunoreactive TSH has reduced bioactivity,105 and in hyperprolactinaemia there may be biologically inactive but immunochemically reactive macro-PRL present in serum.56 Some suspected heterophile antibodies may mimic interference, e.g., elevated hCG in renal disease, menopause, or in the blood of non-pregnant women on haemodialysis.107 In contrast, two-site IMAs that use monoclonal antibodies may result in assay hypersensitivity and a lack of recognition of all hormone isoforms including the biologically active, clinically important forms. Preissner et al.99 recently reported a high heterophile antibody interference rate of >1.5% for a new commercial Tg assay. Masters AM, Hhnel R. Investigation of sex-hormone binding globulin interference in direct radioimmunoassays for testosterone and estradiol. Allergy cross-reactivity occurs when the body identifies allergens as being similar. The highest percentage of heterophile false-positive results in this survey occurred for plasma myoglobin (48% of analyses performed in two of seven tested assay systems). For example, steroids can bind to sex hormone binding globulin36,37 or cortisol, to cortisol binding globulin38 and cause decreased free analyte concentration. Anti-A1 antibodies are considered clinically insignificant unless they react at 37C (99F). [7]:595 Agglutination is scored from 1+ to 4+ based on the strength of the reaction. [28], Donor blood is generally screened for transfusion-transmitted infections such as HIV. Interferences in immunoassay - still a threat [editorial], Leape LL. Effect of anticoagulants and storage temperatures on stability of plasma and serum hormones. Inclusion in an NLM database does not imply endorsement of, or agreement with, Lewis JH, Dusci L, Hackett P, Potter JM. Interfering, endogenous substances that are natural, polyreactive antibodies or autoantibodies (heterophiles), or human anti-animal antibodies together with other unsuspected binding proteins that are unique to the individual, can interfere with the reaction between analyte and reagent antibodies in immunoassay. Drug-specific Fab therapy in drug overdose. [11] The American Society of Hematology recommends that people with sickle cell disease have their blood typed for the RhC/c, RhE/e, Kell, Duffy, Kidd, and MNS antigens prior to transfusion,[12]:1301 because they often require transfusions and may become sensitized to these antigens if transfused with mismatched blood. Martel J, Desprs N, Ahnadi CE, et al. The slide is tilted to mix the cells and reagents together and then observed for agglutination, which indicates a positive result. The test sample is added and the microplate is centrifuged; in a positive reaction, the red blood cells adhere to the surface of the well. Bouvet J-P, Quan CP, Dighiero G. Polyreactivity is not an artefact [letter]. Some antibody screening and identification protocols incorporate a phase of testing after incubation at room temperature, but this is often omitted because most unexpected antibodies that react at room temperature are clinically insignificant. If the test result is indeed negative, the check cells should react with the unbound anti-human globulin and demonstrate agglutination. Ismail AAA, Walker PL, Cawood ML, Barth JH. Interference in immunoassay may lead to the misinterpretation of a patient's results by the laboratory and the wrong course of treatment being given by the physician. The https:// ensures that you are connecting to the Immunometric assay interference: incidence and prevention. The tubes are incubated at body temperature for a defined period of time, then centrifuged and examined for agglutination or hemolysis; first immediately following the incubation period, and then after washing and addition of anti-human globulin reagent. A blood testtogether with an allergy-focused medical history . Rouleaux will disappear once the plasma is replaced with saline, but true agglutination will persist. An unexpected difference between the two results is termed an ABO discrepancy, and must be resolved before the person's blood type is reported. Because the immunoassay test measures the connection of antibodies to a specific corresponding antigen, any connection by the antibody to a similar compound will . . In crossmatching, a prospective transfusion recipient's plasma is added to the donor red blood cells and observed for agglutination (or hemolysis) to detect antibodies that could cause transfusion reactions. They include compounds with chemical differences but structural similarities that cross-react with the antibody, heterophile antibodies, human anti-animal antibodies, autoanalyte antibodies, rheumatoid factors and other proteins. The intricacies of human antibody-mediated HuNoV cross-reactivity and neutralization remain to be fully elucidated. LOUISVILLE, Ky. (WAVE) - The American Red Cross announced the return of its "Stars, Stipes & Pints" Holiday Hero blood drive in Louisville following the Fourth of July holiday. Incidence and mechanism of spurious increases in serum thyrotropin. Other subgroups of A exist, as well as subgroups of B, but they are rarely encountered. Cross-reactivity may be caused by identical carbohydrate structures on unrelated proteins from the same or different species. [7]:590 These analyzers pipette red blood cells and plasma onto gel cards, centrifuge them, and scan and read the agglutination reactions to determine the blood type. Here we report a case of pathological cold agglutinin disease which was eventually detected due to blood group discrepancy. In an example of a case of false hyperprolactinaemia, a number of these procedures were used to identify the presence of a heterophile antibody interference (Table 8).13 The use of three alternate PRL immunoassays indicated a normal PRL value, in keeping with the patients clinical status. [14] Blood group systems other than ABO and Rh have a relatively small risk of complications when blood is mixed, so in emergencies such as major hemorrhage, the urgency of transfusion can exceed the need for compatibility testing against other blood group systems (and potentially Rh as well). Examples of some of the affected analytes are shown in Table 2.98 Bloods from nine of the ten donors resulted in false-positive results of uncertain etiology for six of seven estradiol assay systems (58% of analyses performed) and for two of eight cortisol systems (20% of analyses). Preissner CM, O'Kane DJ, Singh RJ, Morris JC, Grebe SKG. This has led to the suggestion that discordance between IMA and RIA results indicates the presence of assay interference from TgAb.61, The presence of anti-prolactin autoantibodies in the form of macroprolactin (macro-PRL) can cause hyperprolactinaemia without pituitary disease and may lead to unnecessary medical or surgical procedures.56 Macro-PRL is primarily a macro-molecular complex of prolactin (PRL) and an IgG antibody directed against specific epitope(s) on the PRL molecule, and generally is regarded as biologically inactive because of its decreased bioavailability.62 Smith et al.63 report an incidence of 24% of hyperprolactinaemic sera containing macro-PRL with differences of 2.3- to 7.8-fold in PRL concentration observed in ten of these sera using nine immunoassay systems. One-stage technique. The immunoassay handbook. This method is typically used in under-resourced areas or emergency situations; otherwise, alternative methods are preferred. [20], Crossmatching, which is routinely performed before a blood transfusion, involves adding the recipient's blood plasma to a sample of the donor's red blood cells. [9]:21011, Serologic methods for blood compatibility testing make use of these antibody-antigen reactions. By Dustin Vogt. Hursting MJ, Raisys VA, Opheim KE. If a clinically significant antibody is detected (or was in the past), or if the immediate spin crossmatch demonstrates incompatibility, a "full" or "IgG crossmatch" is performed, which uses the indirect antiglobulin test to detect blood group incompatibility caused by IgG antibodies. 8600 Rockville Pike Bethesda, MD 20894, Web Policies The recognition of such aberrant test results requires constant surveillance by both laboratory and physician. A diagrammatic representation of the elimination of interference from antibody-binding substances (I) binding to the reagent antibodies in two-site immunoassays by the addition of non-human immunoglobulins (Ig), which binds to I. Reproduced with the permission of Clinical Chemistry from Ref 79. [1]:261 Genetic testing is required for typing red blood cell antigens for which no commercial antisera are available. Warm autoimmune hemolytic anemia, where antibodies react . Cross-reacting species also result in over- or underestimation of sample analyte concentration if an immunoassay reagent contains antibodies directed toward molecules other than the antigen of interest. An adaptive immune response is specific to the antigen that stimulated it (called the immunogen). [9]:214 Microplate wells coated with anti-A, -B and -D reagents are used for forward grouping. If the blood is incompatible, the antibodies in the recipient's plasma will bind to antigens on the donor red blood cells. official website and that any information you provide is encrypted Reducing interference from heterophilic antibodies in a two-site immunoassay for carcinoembyronic antigen (CEA) by using a human/mouse chimeric antibody to CEA as the tracer. Cells that have not agglutinated collect on the bottom. The interfering antibody substances in immunoassay have several properties in common (Table 3). Another procedure for detecting and identifying a suspected interfering antibody is measurement before and after the addition of a blocking reagent to the sample and the finding of a significantly different result (Table 7). (Wash.) , 90 ( 1993 ) , pp. Want to thank TFD for its existence? Cross-reactivity, in a general sense, is the reactivity of an observed agent which initiates reactions outside the main reaction expected. Tissue cross-reactivity assay is a standard method based on immunohistochemistry, required prior to phase I human studies for therapeutic antibodies. An example of confounding that yields a false positive error is in a latex fixation test when agglutination occurs with another antigen rather than the antigen of interest. Analytical performance of free and total thyroxine assays. . He observed that a person's serum never agglutinated their own red blood cells, but it could agglutinate others', and based on the agglutination reactions the red cells could be sorted into three groups: group A, group B, and group C. Group C, which consisted of red blood cells that did not react with any person's plasma, would later be known as group O. Steimer W, Mller C, Eber B. Digoxin assays: frequent, substantial, and potentially dangerous interference by spironolactone, canrenone, and other steroids. Interferences that alter the measurable analyte concentration in the sample, Pre-analytical factors, e.g., anticoagulants, sample storage, Interferences that alter antibody binding, Interfering antibody concentration can change over time within the same individual, Low affinity polyspecific antibodies can be present in high concentrations or high affinity in low concentrations, Can produce falsely high (false-positive) or falsely low (false-negative) results, May interfere within one or more manufacturers' immunoassay systems but not necessarily in all assays, May interfere in a number of immunoassays for different analytes, The inclusion of one or more interference blocking agents in manufacturers' immunoassay reagents may be insufficient to overcome the interference, Prior extraction of analyte from sample, e.g., by chromatography, - Murine or other animal species serum immobilised onto Sepharose beads, Polyethylene glycol precipitation (PEG 6000), Heating to 7090C (for heat-stable analytes only), Addition of blocking agent from the same species as the antibody reagents, Non-immune serum, species-specific polyclonal IgG, anti-human IgG or polymerised mouse IgG, Species-specific fragments of IgG [Fc, Fab, F(ab'), Heterophile blocking reagents (HBR), immunoglobulin inhibiting reagent (IIR), and antibody blocking tubes, Use of an alternate immunoassay that preferably uses antibody raised to a different species, Measurement before and after addition of a blocking reagent, especially bovine, or a series of concentrations of the blocker, or a combination of blockers from different species, Measurement of dilutions of the sample using the manufacturer's diluent containing non-immune globulin, Radioimmunoprecipitation of labelled thyroid hormones to detect anti-T3 and anti-T4 autoantibodies. Routine blood typing involves determining the ABO and RhD (Rh factor) type,[note 1] and involves both identification of ABO antigens on red blood cells (forward grouping) and identification of ABO antibodies in the plasma (reverse grouping). The prevalence of immunoassay interference is variable and dependent on the type of antibody interference. The strength of positive reactions is scored from 1+ to 4+ depending on how far the cells have travelled through the gel. Heterophiles can bind to the conjugate, enzyme, or other parts of the detection system in reagent-limited assays, mainly of the non-RIA type, and cause interference in assays for steroid hormones, thyroid function tests, and digoxin.7,7678 In conventional two-site IMAs for the measurement of non-immunoglobulin antigen, heterophile antibody or any multivalent antibody-binding substance binds animal capture antibody to the detection antibody and falsely elevates the patient value by producing an assay signal (Figure 1).79 Assays using either polyclonal or monoclonal antibodies may be affected. Weak reactions in forward grouping can be strengthened by incubating the blood and reagent mixture at room temperature or 4C (39F), or by using certain enzymes to enhance the antigen-antibody reactions. The sample properties are unique to the patient and interference results from an interaction with one or more steps in the immunoassay procedure such that the measurable analyte concentration in the sample or antibody binding is altered (Table 1).19,20 Other unsuspected binding protein(s) in the individual also can cause interference in immunoassay by interfering with the reaction between analyte and assay antibodies. Levinson SS, Miller JJ. Blood compatibility testing is routinely performed before a blood transfusion. Cross-reactions are common in blood tests for fluke infections because the different species are closely related. Russians spilling Russian blood. Wharton's jelly can be removed by thoroughly washing the red blood cells. Hunter WM, Budd PS. FOIA At the same time senior staff should be proactive in improving the laboratory-clinical communication link by presentation and discussion of laboratory data at local journal club meetings, etc. Crossmatching is a test used to check for harmful interactions between your blood and specific donor blood or organs. Spencer CA, Takeuchi M, Kazarosyan M, et al. Samples or assay reagents contaminated with substances that interfere with measurement of the label, e.g., enzyme inhibitors, fluorophores from ophthalmic examination, or radioimaging isotopes, require removal by washing. It is equally important that physicians communicate any clinical suspicion of discordance between the clinical and the laboratory data to the laboratory. Cross-reaction is a problem in diagnostic immunoassays where endogenous molecules with a similar structure to the measured analyte exist or where metabolites of the analyte have common cross-reactive epitopes, and where there is administration of structurally similar medications.2 Early hCG immunoassays were cross-reactive with luteinizing hormone (LH)25 but the development of more specific antibodies has led to most of todays assays for hCG having little or no cross-reaction with LH. A + means that the antigen is present on the reference red blood cells, and 0 means it is absent; nt means "not tested". The physiological condition of the patient can lead to misinterpretation of results due to differences in metabolism, receptor binding, or post-translational defects, e.g., the sick euthyroid patient. Gagajewski A, Davis GK, Kloss J, Poch GK, Anderson CJ, Apple FS. Materials and Methods: In this cross-sectional study, data of blood donors were obtained from the integrated database of Yazd Blood Transfusion Center during a period of eight . The invention of the antiglobulin test led to the discovery of many more blood group antigens. Jones G. Handling common laboratory interferences. [1]:161 Antibodies that are predominantly IgG, such as those directed towards antigens of the Duffy and Kidd systems,[1]:198200 generally do not cause immediate agglutination because the small size of the IgG antibody prevents formation of a lattice structure. *For correspondence: Ms Jill Tate e-mail: The contents of articles or advertisements in The Clinical Biochemist Reviews are not to be construed as official statements, evaluations or endorsements by the AACB, its official bodies or its agents. Steimer W. Performance and specificity of monoclonal immunoassays for cyclosporine monitoring: how specific is specific? If the result is caused by an alloantibody, an antibody screen can be performed to identify the antibody,[7]:1412 and the reverse grouping can be performed using samples that lack the relevant antigen. Fiad TM, Duffy J, McKenna TJ. By the early 1950s, companies had begun producing commercial antisera for special antigen testing. The interference may result in discordant results for one or more analytes, and may be detected in one or more other assay systems for the affected analyte. Kuroki M, Matsumoto Y, Arakawa F, et al. On July 5 and 6, the blood drive will be hosted at the UAW Local 862s Union Hall at 3000 Fern Valley Road . There are four major blood groups determined by the presence or absence of two antigens, A and B, on the surface of red blood cells. [3] Cross-reactivity need not be between closely related viruses, however; for example, there is cross-reactivity between influenza virus-specific CD8+ T cells and hepatitis C virus antigens. President Vladimir Putin calls the armed rebellion by the Wagner Group chief Yevgeny Prigoshin a "betrayal." . If this is not effective, dithiothreitol can be used to destroy the antibodies. "Blood type" refers to a specific pattern of reaction to testing antisera within a given system. Limitations of the radioimmunoassay for Hepatitis B antigen [letter]. Ismail AAA, Walker PL, Barth JH, Lewandowski KC, Jones R, Burr WA. Prince AM, Brotman B, Jass D, Ikram H. Specificity of the direct solid-phase radioimmunoassay for detection of the Hepatitis-B antigen. An example of helpful cross-reactivity is in heterophile antibody tests, which detect Epstein-Barr virus using antibodies with specificity for other antigens. Usually, when a food . [7]:6003 In emergencies, blood may be issued before crossmatching is complete. Multicenter study of Abbott AxSYM. For these reasons there may be a higher prevalence of unpredictable cross-reaction in IMAs compared with a single-site antigen-antibody reaction in reagent-limited assays.21 Exogenous antibodies given to a patient for therapy may also compete with the assay antibody for the analyte and disturb the antigen-antibody reaction resulting in immunoassay interference, e.g., administration of Fab fragments derived from anti-digoxin antibodies (Digibind).22. Therefore, blood typing using IgG antisera and detection of IgG antibodies requires use of the indirect antiglobulin test to demonstrate IgG bound to red blood cells.
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