Epstein-Barr病毒壳抗原gp125的纯化和应用

目的研制ｅｐｓｔｅｉｎ－Ｂａｒ（ＥＢ）病毒诊断试剂。方法将重组痘苗病毒表达的Ｅｐｓｔｅｉｎ－Ｂａｒ病毒（ＥＢＶ）壳抗原（ＶＣＡ）主要多肽ｇｐ１２５纯化，作为诊断抗原建立了酶联免疫吸附试验（ＥＬＩＳＡ），检测了４８份鼻咽癌（ＮＰＣ）病人血清及１０份正常人血清中的ＶＣＡ／ＩｇＡ抗体。结果该方法与免疫荧光（ＩＦ）检测结果一致，但ＥＬＩＳＡ的平均几何滴度（ＧＭＴ）是ＩＦ的１２倍。结论以纯化的ＥＢ病毒壳抗原主要多肽ｇｐ１２５作为诊断抗原建立的检测方法，更适合于ＥＢＶ相关疾病的血清学诊断和血清流行病学调查。     

Contribution to rapid diagnosis of infectious mononucleosis

By indirect immunofluorescence (IF) technique humoral antibodies to Epstein-Barr virus capsid antigen (EB-VCA) and to cytomegalovirus (CMV) were detected in 47% and 9% of persons with infectious mononucleosis (IM), respectively. In 23% of the patients examined, IgM antibodies to both viruses were detected, while in 8% of them high titres of IgG only were found in the absence of IgM class antibodies to EB-VCA or to CMV. The finding of IgM antibody to EB-VCA was in good correlation with the persisting symptoms of the disease. Discrepancy between the presence of specific IgM and the absence of heterophilic antibodies was observed in some children and in all persons with persistent or recurrent signs of IM. In the latter, specific IgM was found only during exacerbation of the disease, but during remissions IgG antibodies persisted in high levels. Antibodies to Epstein-Barr virus nuclear antigen (EBNA) were detected in all chronically ill persons and antibodies to the R-component of Epstein-Barr virus early antigen (EA) were present in the majority of them.     

Enhancement of Epstein-Barr virus antibody production in systemic lupus erythematosus patients.

Two groups of 22 patients suffering from either systemic lupus erythematosus (SLE) or infectious mononucleosis (IM) were checked for Epstein-Barr virus capsid antigen antibody (EB-VCA) production. The average significant antibody levels as well as the frequency of their occurrence were clearly higher in SLE than in IM patients.     

A novel EBV-related nucleo-cytoplasmic antigen in a null cell-line (HLN-STL-C) reactive to antibodies in the sera from patients with Sj?gren's syndrome

We have established a non-T- and non-B-cell line, HLN-STL-C(STL-C), which harbors the EBV genome, from the lymph node cells of a Japanese ATL patient. This cell line expresses a unique Epstein-Barr virus (EBV)-related nucleo-cytoplasmic (N-C) antigen which is detected by indirect immunofluorescence (IF) with the sera from patients with nasopharyngeal cancer (NPC), infectious mononucleosis (IM) or adult T-cell leukemia (ATL). One of the molecular components of this antigen is proved to be STL-C specific 125 kD molecule by immunoblot analysis (IB). To study the involvement of EBV in Sj?gren's syndrome (SS), we examined the reactivity of the N-C antigen with the sera of SS patients by IF and IB. Among 24 cases examined, the sera of 21 cases (87.5%) positively stained the N-C antigen by IF. The staining patterns were divided into two types. Type I, (seven cases) showed positive staining for only N-C antigen, and Type II, (14 cases) was positive for N-C antigen associated with diffuse nuclear staining due to antinuclear antibodies in the SS patient's sera. Only one out of 11 non-Sj?gren's patients' sera, which were almost all healthy controls, was positive for N-C antigen in this study. By IB, however, only two out of 15 IF-positive SS patients' sera reacted with STL-C specific 125 kD molecule. These results suggested the presence of heterogenous components in the N-C antigen. Our findings may support the hypothetical conception that EBV plays an etiological role in SS.     

Development of an antibody-capture IgM-enzyme-linked immunosorbent assay for diagnosis of acute Epstein-Barr virus infections

An anti-EBV IgM-ELISA was developed using the antibody-capture principle, to be used for the diagnosis of acute infectious mononucleosis (IM). The test was based on anti-human IgM-coated microtiter plates; nuclei of EBV producer cells were used for antigen; conjugate was prepared by labeling sheep anti-EBV IgG with horseradish peroxidase. The specificity of the anti-EBV IgM-ELISA was studied with a panel of sera from acute infections with hepatitis A virus, rubella virus, Toxoplasma gondii and cytomegalovirus, and sera positive for rheumatoid factors, positive for antinuclear antibodies, as well as with sera from normal blood donors and pregnant women. Specificity in these panels was 98.4%. In a clinical study with 449 sera from patients with IM-like symptoms, 109 of 109 confirmed patients were detected by the anti-EBV IgM-ELISA. Specificity of the anti-EBV IgM-ELISA in this clinical study was 99.7%. The anti-EBV IgM-ELISA detected several acute EBV patients who had negative heterophile antibody titers.     

Antiviral neutralizing antibody to Hantaan virus as determined by plaque reduction technique

Summary The 76–118 strain of Hantaan virus was titrated in E6 (Vero) cells by the plaque method using agarose overlay medium. Visible plaques, formed 10 days post-infection, were uniformly 2–3 mm in diameter. Dose-response experiments showed that a single infectious particle initiated the formation of a plaque. Infectivity titers by the plaque method were equivalent to those obtained by the endpoint method (TCID₅₀) using the immunofluorescence antibody technique (IFA) for antigen detection. The single-cycle growth pattern of the virus showed an eclipse phase of 7 to 9 hours, with production of cell-free infectious virus 18 hours post-infection. Plaque reduction neutralization tests suggested that complement enhanced the neutralizing activity of sera; rat sera were particularly complement-dependent. The plaque reduction neutralization test was about 10 times more sensitive than the TCID₅₀ neutralization test. Convalescent phase sera from patients with hemorrhagic fever with renal syndrome (HFRS) having higher IF antibody titers to Hantaan virus than to nephropathia epidemica (NE) virus were capable of neutralizing Hantaan virus, while sera from patients with higher IF antibody titers to NE virus than Hantaan virus did not contain neutralizing antibody to Hantaan virus.With 3 FiguresOn leave from the Department of Virology, Kyushu University School of Medicine, Fukuoka 812, Japan.     

Serodiagnosis of infectious mononucleosis by using recombinant Epstein-Barr virus antigens and enzyme-linked immunosorbent assay technology.

Four recombinant, diagnostically useful Epstein-Barr virus (EBV) proteins representative of the viral capsid antigen (p150), diffuse early antigen (p54), the major DNA-binding protein (p138), and the EBV nuclear antigen (p72) (W. Hinderer, H. Nebel-Schickel, H.H. Sonneborn, M. Motz, R. Kühbeck, and H. Wolf, J. Exp. Clin. Cancer Res. 7[Suppl.]:132, 1988) were used to set up individual enzyme-linked immunosorbent assays (ELISAs) for the qualitative and quantitative detection of immunoglobulin M (IgM) and IgG antibodies. In direct comparison with results obtained by standard immunofluorescence or immunoperoxidase assays, it was then shown that the recombinant EBV ELISAs provide the means for specific and sensitive serodiagnosis of infectious mononucleosis (IM) caused by EBV. The most useful markers in sera from such patients proved to be IgM antibodies against p54, p138, and p150. Additional positive markers for recent or ongoing IM apparently were IgG antibodies against p54 and p138. In contrast, anti-p72 IgG had a high preference for sera from healthy blood donors and, therefore, can be considered indicative of past exposure to the virus. Altogether, the individual ELISAs proved to be as specific and at least as sensitive for the diagnosis of IM as the currently available standard techniques are. Moreover, our findings suggest that, by combining individual test antigens, a workable ELISA system consisting of three assays (IgM against p54, p138, and p150; IgG against p54 and p138; and IgG against p72) can be established for the standardized rapid diagnosis of acute EBV infections.     

Limitations of available tests for diagnosis of infectious mononucleosis.

Among 500 students seen at a university health service for illnesses resembling infectious mononucleosis (IM), the diagnosis of IM was established in 124 (25%) on the basis of the initial presence or subsequent emergence of the spectrum of Epstein-Barr virus (EBV) antibodies characteristic of a primary EBV infection. Of these 124 patients, 113 had an EBV-specific antibody pattern in the initial serum indicative of current primary infection; however, 11 (9%) had no detectable immunoglobulin G antibodies to EBV-specific antigens in their first serum. The sensitivity of this panel of EBV antibody assays was 91% and the specificity was 100%. Initial sera had detectable heterophil antibodies for 107 (86%) of the 124 students with IM and for 2 with other illnesses. Among our patients, the Monospot (Ortho Diagnostics Inc.) test had a sensitivity of 86% and a specificity of 99%. Reliance on hematological criteria (lymphocyte count greater than or equal to 50% and atypical lymphocyte count greater than or equal to 10%) gave a sensitivity of only 39%, but a specificity of 99%. Students with IM who showed a delayed emergence of the spectrum of EBV-specific antibodies characteristic of an acute infection were compared with control patients who had such antibodies at the time of their initial visit to the health service. They were found to have a briefer duration of illness (P greater than 0.05), lower leukocyte (P less than 0.005), lymphocyte (P less than 0.005), and atypical lymphocyte (P less than 0.05) counts, and a less frequent occurrence of heterophil antibodies (P less than 0.05).     

Epstein-Barr virus serology in bone marrow transplantations: a one-year retrospective study with detection of EBV IgM-VCA-specific antibodies

The specific antibody response to Epstein-Barr virus (EBV) antigens of 41 bone marrow transplant recipients with leukemia or aplastic anemia was examined retrospectively by immunofluorescence test (IF) over 1 year. We observed high titers (greater than 640) of IgG-viral capsid antigen (VCA) with emergence of IgG-early antigen (EA) and frequent absence or low levels of Epstein-Barr nuclear antigen (EBNA) antibodies. After absorption to remove rheumatoid factor (RF), five of the 41 recipients had IgM-VCA antibody to EBV, which appeared between weeks 26 and 48 after BMT and persisted for 1-4 months. No heterophil antibodies were detected in these sera, and none of the five recipients had a history of infectious mononucleosis.     

Antibodies to Epstein-Barr virus in nasopharyngeal carcinoma, tonsillar carcinoma and control groups

In the sera of 17 patients with nasopharyngeal carcinoma (NPC) and of 19 patients with tonsillar carcinoma (TC) the titres of IgA, IgG and IgM antibodies to EBV VCA (viral capsid antigen) and of IgG antibodies to EBV EA (early antigen) were determined by the indirect immunofluorescence (IF) method. Significant difference was observed in the frequency of IgA antibodies to EBV VCA and IgG antibodies to EBV EA between NPC patients and controls. There was also a significant difference between the frequency of IgM antibody to EBV VCA and EBV EA antibody titres in TC patients and controls. The geometric mean titre (GMT) of IgG antibodies to EBV VCA was significantly higher in the NPC and TC patients as compared to controls.     

Detection of IgA antibody to EBV membrane antigen using Staphylococcus aureus preabsorbed sera is closely associated with nasopharyngeal carcinoma

The presence of IgA antibody to membrane antigen (MA) of Epstein-Barr virus (EBV) was tested in sera from 48 nasopharyngeal carcinoma (NPC) patients, 40 patients with tumors other than NPC and 46 normal individuals. The sera were preabsorbed with Staphylococcus aureus (SPA) (strain no. 1800) prior to their use in the indirect immunofluorescence test. One hundred percent of the NPC patients had the IgA/MA antibody with a GMT of 1:141. In patients with tumors other than NPC or normal individuals, IgA/MA antibodies were not detectable. The IgA/MA antibodies have been demonstrated in 6 NPC patients lacking detectable antibody levels in the indirect immunofluorescence test using nonabsorbed sera. Our data indicate that preabsorbtion of sera with SPA renders the diagnostic test significantly more sensitive for the detection of the nasopharyngeal carcinoma and can be used for trials on the prognosis of patients.     

A human monoclonal autoantibody isolated from a patient with infectious mononucleosis reactive with both self antigens and Epstein-Barr virus nuclear antigen (EBNA)

In order to investigate the mechanism(s) by which Epstein-Barr virus (EBV) induces the outcome of autoantibodies during infectious mononucleosis (IM), a human IgM (k) monoclonal antibody to cytoskeletal filaments of epithelial cells has been prepared by EBV transformation of peripheral blood B lymphocytes obtained from a patient with IM. The antibody was also found to react with smooth muscle of frozen sections of human stomach tissue by immunofluorescence, and with the Epstein-Barr nuclear antigen (EBNA) by an enzyme-linked immunosorbent assay. These findings demonstrate at the clonal level the epitope homology between host's cell antigens and EBV-encoded nuclear antigen, which might have relevance in EBV-induced autoimmunity.     

Detection of an idiotope on a human monoclonal autoantibody by monoclonal anti-idiotypic antibody and its relationship to Epstein-Barr virus-induced autoimmunity.

We have recently described a human IgM monoclonal antibody (mAb), reactive with both self antigens, i.e., cytoskeleton filaments and smooth muscle, and Epstein-Barr virus (EBV)-induced nuclear antigen (EBNA), produced by EBV-transformed B lymphocytes isolated from a patient with infectious mononucleosis (IM). In order to achieve higher antibody secretion in culture supernatant, the mAb-producer cells were fused with ouabain-resistant mouse myeloma cells and a stable human-mouse heterohybrid, coded HY 5488, producing up to 80 micrograms/ml IgM mAb, was isolated after 4 cloning procedures. Purified HY 5844 mAb was used to immunize mice for the production of a murine anti-idiotypic mAb, which was used to probe the expression of the idiotope of HY 5488 mAb (Id 5488) in sera of IM patients and normal controls by ELISA. It was found that Id 5488 is expressed both in IM patients and normal controls, and that Id 5488 expression is significantly higher in IM patients' sera; furthermore, in IM sera a statistically significant correlation between Id 5488 expression and anti-cytoskeleton and anti-smooth muscle autoantibodies was found. It is suggested that at least part of EBV-induced IgM autoantibodies appearing during IM are secreted by B lymphocytes programmed to the production of "natural antibodies" bearing Id 5488-like idiotopes.     

The response to Epstein-Barr virus infection in Sj?gren's syndrome

To assess the response to Epstein-Barr virus (EBV) infection in patients with primary Sj?gren's syndrome (SS), the frequency of detection of EBV DNA was studied in salivary gland biopsies and the antibody and idiotypic response to the virus was compared with healthy controls and infectious mononucleosis (IM). Viral DNA, detected by in-situ hybridization, was found in biopsies from two out of 12 patients with SS and six out of 10 controls. IgG, IgA and IgM antibodies to the virus, measured by ELISA using synthetic peptides (early antigen and EBNA-1) and a cloned fusion protein (EBNA-1), were normal in sera from 20 patients with SS, whereas infectious mononucleosis patients showed an increase in IgM antibodies to EBNA-1 and IgG antibodies to early antigen. One similarity between infectious mononucleosis and Sj?gren's syndrome was a significant increase in the germline heavy chain idiotype G6 in both diseases, suggesting activation of similar B-cell subsets. It is possible that this is due to EBV, though the low frequency of EBV DNA in biopsies and the normal levels of EBV antibodies in SS does not lend any evidence that the virus itself is the causative agent.     

Identification of an Epstein-Barr virus early gene encoding a second component of the restricted early antigen complex

When the latent Epstein-Barr virus (EBV) genome in B95-8 cells is induced into a replicative phase, two abundant early RNAs are transcribed rightward from the EBV BamHI H DNA fragment into BamHI F. Analysis of cDNA clones prepared from the RNA of cells replicating EBV revealed that both RNAs contain the BHRF1 open reading frame. Part of BHRF1, cloned into a prokaryotic fusion protein expression vector, expressed a fusion protein in Escherichia coli and the purified fusion protein was used to generate a monoclonal antibody against BHRF1. This antibody was then employed to characterize the protein encoded by BHRF1 in cells replicating EBV. The monoclonal antibody reacted with a 17-kDa protein component of the restricted early antigen (EA) complex. The distribution of the protein in cells was similar to that noted when sera from patients with African Burkitt's lymphoma were used to stain these cells. The protein was synthesized before the major 47-56 kDa protein associated with the diffuse component of EA in superinfected Raji cells. All human sera containing antibodies to EA as determined by immunofluorescence (IF) reacted with the protein as did some sera determined to be anti-VCA positive and anti-EA negative by IF. The predicted amino acid sequence of the protein has characteristics which suggest that it is a membrane protein. It also has significant homology with both the anchor region of polyoma middle T antigen and with the predicted protein product of the bcl-2 mRNA activated by the 14/18 chromosome translocation characteristic of follicular lymphomas. This latter homology is extensive and colinear, suggesting common evolution and function. However, neither a mRNA which could efficiently translate the BHRF1 protein nor the BHRF1 protein could be detected in latently infected cells. Thus, the bcl-2 predicted protein is similar to an EBV protein synthesized in the early phase of virus infection.     

Evaluation of an Immunofiltration Assay That Detects Immunoglobulin M Antibodies against the ZEBRA Protein for the Diagnosis of Epstein-Barr Virus Infectious Mononucleosis in Immunocompetent Patients

The performance of an immunofiltration assay (IMFA) that detects immunoglobulin M (IgM) antibodies to the Epstein-Barr virus (EBV) ZEBRA (BamHI Z EBV replication activator) protein was evaluated for the diagnosis of EBV infectious mononucleosis (IM) in immunocompetent patients. The test panel consisted of 47 sera displaying an EBV-specific antibody profile compatible with an acute primary EBV infection from patients with clinical and biological features of EBV IM, 20 sera from healthy individuals either with a past EBV infection or who were EBV seronegative, 20 sera displaying an equivocal EBV antibody pattern (viral capsid antigen IgG positive [VCA IgG⁺], VCA IgM⁺, and EBV nuclear antigen-1 IgG⁺), and 15 sera obtained from patients with a mononucleosis-like syndrome owing to cytomegalovirus, human herpesvirus 6, or parvovirus B19. Overall, the sensitivity and the specificity of the assay were found to be 92.5%, and 97.3%, respectively. The sensitivity of the assay for the diagnosis of heterophile antibody-negative EBV IM was 86.2%. The IMFA is rapid, easy to perform, and, thus, suitable for point-of-care testing, and it may be used as a first-line test for the diagnosis of acute EBV IM in immunocompetent patients.Diagnosis of Epstein-Barr virus (EBV) infectious mononucleosis (IM) is commonly made on the basis of characteristic clinical manifestations and the detection of heterophile antibodies (HA). Nevertheless, HA may be absent, particularly in young children (14) but also in as many as 20% of adults with EBV IM (7). In these cases, demonstration of the presence of EBV viral capsid antigen (VCA) immunoglobulin G (IgG) and/or IgM antibodies, along with the absence of IgG antibodies to EBV nuclear antigen-1 (EBNA-1), allows the diagnosis of EBV primary infection (9). Detection of EBV-specific antibodies is accomplished by the use of commercial enzyme immunoassays, indirect immunofluorescence assays, line blot immunoassays (9), or, as established more recently, a multiplexed bead assay (3). These methods have long turnaround times, are labor-intensive, or require specific instruments or skilled technologists for their performance. In addition, interpretation of EBV VCA IgG/IgM and EBNA-1 IgG reactivity profiles is not always straightforward (9).The ZEBRA (BamHI Z EBV replication activator) protein is encoded by the immediate early BZLF1 gene. ZEBRA is expressed during the lytic cycle in EBV-permissive cells and plays a critical role in transactivating several immediate early, early, and late EBV genes (5). Antibodies against ZEBRA are produced during primary EBV infection (11, 15, 18), and thus, the detection of ZEBRA-specific IgMs may allow an early diagnosis of EBV IM. In the present study, we evaluated a rapid and easy-to-perform immunofiltration assay (IMFA) detecting IgMs to the EBV ZEBRA protein for the biological diagnosis of IM in immunocompetent patients.     

Evaluation of 12 commercial tests for detection of Epstein-Barr virus-specific and heterophile antibodies

Ten microbiological departments in Norway have participated in a multicenter evaluation of the following commercial tests for detection of Epstein-Barr virus (EBV)-specific and heterophile antibodies: CAPTIA Select viral capsid antigen (VCA)-M/G/EBNA (Centocor Inc.), Enzygnost anti-EBV/immunoglobulin M (IgM) and IgG (Dade Behring), Vironostika EBV VCA IgM/IgG/EBNA enzyme-linked immunosorbent assay (ELISA) (Organon Teknika), SEROFLUOR immunofluorescence assay and EBV Combi-Test (Institute Virion Ltd.), anti-EBV recombinant IgM- and IgG-early antigen/EBNA IgG ELISA (Biotest Diagnostics), EBV IgM/IgG/EBNA ELISA (Gull Laboratories), Paul-Bunnell-Davidsohn test (Sanofi Diagnostics Pasteur), Monosticon Dri-Dot (Organon Teknika), Avitex-IM (Omega Diagnostics Ltd.), Alexon Serascan infectious mononucleosis test (Alexon Biomedical Inc. ), Clearview IM (Unipath Ltd.), and Cards+/-OS Mono (Pacific Biotech, Inc.). The test panel included sera from patients with primary EBV infection, immunocompromised patients with recent cytomegalovirus infection, healthy persons (blood donors), and EBV-seronegative persons. Among the tests for EBV-specific antibodies the sensitivity was good, with only small differences between the different assays. However, there was a greater variation in specificity, which varied between 100% (Enzygnost) and 86% (Biotest). Tests for detection of heterophile antibodies based on purified or selected antigen (Avitex, Alexon, Clearview IM, and Cards+/-OS Mono) were more sensitive than the Paul-Bunnell-Davidsohn and Monosticon tests.     

Radioimmunoprecipitation in the diagnosis of chronic active epstein-barr virus infection

Chronic active Epstein-Barr virus (EBV) infection occurs sporadically in a small fraction of individuals infected with EBV. A clear definition of the disease and an unambiguous diagnostic test are still lacking. In an attempt to identify a serologic marker to facilitate the diagnosis, immunoblot and radioimmunoprecipitation assay (RIPA) were compared with standard immunofluorescence on 39 available sera. Results by RIPA revealed that antibodies to a 120 kDa viral protein correlated with the presence of chronic active EBV infection; these antibodies were not detected in sera from other EBV-seropositive individuals, with the exception of one of two patients with ataxia telangiectasia. Also, RIPA was the most sensitive technique for detecting EBV antibodies in sera weakly or doubtfully positive for antibody to EB viral capsid antigen by indirect immunofluorescence. All these sera had antibodies to the 150 kDa protein, also known as p160, the major viral capsid antigen. © 1994 Wiley-Liss, Inc.     

Complement-Fixing Antigens in Burkitt Lymphoma Cell Lines

The complement-fixing (CF) activity of antigens from cultured Burkitt lymphoma cells was determined by using normal American sera as the source of antibody. Approximately 75% of the sera fixed complement with the positive cell lines. These lines contained the herpes-like virus detectable by electron microscopy. The content of CF antigen depended on the cell line used but appeared to be independent of the number of cells which produced Henle's immunofluorescence (IF) antigen. Only sera that reacted in the IF test also contained CF antibodies to the crude cell extracts.     

Evaluation of 12 Commercial Tests for Detection of Epstein-Barr Virus-Specific and Heterophile Antibodies

Ten microbiological departments in Norway have participated in a multicenter evaluation of the following commercial tests for detection of Epstein-Barr virus (EBV)-specific and heterophile antibodies: CAPTIA Select viral capsid antigen (VCA)-M/G/EBNA (Centocor Inc.), Enzygnost anti-EBV/immunoglobulin M (IgM) and IgG (Dade Behring), Vironostika EBV VCA IgM/IgG/EBNA enzyme-linked immunosorbent assay (ELISA) (Organon Teknika), SEROFLUOR immunofluorescence assay and EBV Combi-Test (Institute Virion Ltd.), anti-EBV recombinant IgM- and IgG-early antigen/EBNA IgG ELISA (Biotest Diagnostics), EBV IgM/IgG/EBNA ELISA (Gull Laboratories), Paul-Bunnell-Davidsohn test (Sanofi Diagnostics Pasteur), Monosticon Dri-Dot (Organon Teknika), Avitex-IM (Omega Diagnostics Ltd.), Alexon Serascan infectious mononucleosis test (Alexon Biomedical Inc.), Clearview IM (Unipath Ltd.), and Cards±OS Mono (Pacific Biotech, Inc.). The test panel included sera from patients with primary EBV infection, immunocompromised patients with recent cytomegalovirus infection, healthy persons (blood donors), and EBV-seronegative persons. Among the tests for EBV-specific antibodies the sensitivity was good, with only small differences between the different assays. However, there was a greater variation in specificity, which varied between 100% (Enzygnost) and 86% (Biotest). Tests for detection of heterophile antibodies based on purified or selected antigen (Avitex, Alexon, Clearview IM, and Cards±OS Mono) were more sensitive than the Paul-Bunnell-Davidsohn and Monosticon tests.