Immune Adherence Hemagglutination Assay Research Articles

Sensitivity and specificity of different antibody tests for detecting varicella-zoster virus

IntroductionAntibody tests for detecting varicella-zoster virus include the fluorescent-antibody-to-membrane-antigen (FAMA) assay, immune adherence hemagglutination assay (IAHA), enzyme immunoassay (EIA), and the glycoprotein-based enzyme-linked immunosorbent assay (gpELISA). Although FAMA and gpELISA are highly sensitive, FAMA is not available commercially. Therefore, this study was performed to compare potential high-sensitivity tests with commercially available tests. MethodsFour antibody tests, FAMA, gpELISA, EIA, and IAHA, were performed using sera collected from 32 children aged 7 months–10 years. Using FAMA as a reference, the sensitivity and specificity of gpELISA, EIA, and IAHA were assessed. Subsequently, using gpELISA as a reference, the positive agreement rate of EIA and IAHA was assessed. ResultsOn a reference scale with FAMA set at 100%, the sensitivity and specificity of the antibody tests were as follows: gpELISA, 67% and 100%; EIA, 67% and 100%; and IAHA, 47% and 100%, respectively. The positive agreement rates of EIA and IAHA relative to gpELISA were 86% and 64%, respectively.Conclusions: gpELISA had a lower positive rate than did FAMA, and showed comparable sensitivity to that of EIA.

Results of Booster Vaccination in Children with Primary Vaccine Failure after Initial Varicella Vaccination

In October 2014, the varicella vaccination policy in Japan was changed from a single voluntary inoculation to two routine inoculations. This paper reports the results of booster vaccination in children who did not show seroconversion after initial vaccination (i.e., primary vaccine failure : PVF) over a 7-year period prior to the introduction of routine varicella vaccination. Between November 2007 and May 2014, 273 healthy children aged between 1.1 and 14.5 years (median : 1.7 years) underwent varicella vaccination. Before and 4 to 6 weeks after vaccination, the antibody titers were measured using an immune adherence hemagglutination (IAHA) assay and a glycoprotein-based enzyme-linked immunosorbent assay (gpELISA). In addition, side reactions were examined during the four-week period after vaccination. Children who did not show IAHA seroconversion (PVF) were recommended to receive a booster vaccination, and the measurement of antibody titers and an assessment of side reactions were performed after the booster dose. In May 2015, a questionnaire was mailed to each of the 273 participants to investigate whether they had developed varicella and/or herpes zoster after vaccination. After initial vaccination, the IAHA seroconversion rate was 75% and the mean antibody titer (Log2) with seroconversion was 4.7, while the gpELISA seroconversion rate was 84% and the mean antibody titer (Log10) with seroconversion was 2.4. Among children with PVF, 54 received booster vaccination within 81 to 714 days (median : 139 days) after the initial vaccination. After booster vaccination, the IAHA seroconversion rate was 98% and the mean antibody titer (Log2) with seroconversion was 5.8. Both the seroconversion rate and the antibody titer were higher compared with the values after the initial vaccination (p < 0.01). After booster vaccination, the gpELISA seropositive rate was 100% and the mean positive antibody titer (Log 10) was 3.6 ; similar results were obtained for the IAHA assay, with a significantly higher, antibody response than that after the initial vaccination (p < 0.01). Side reactions were generally minor, including fever (≥ 37.5 degrees C), rash at the injection site, and rash at other sites. There were no significant differences in the incidences of side reactions between the initial and booster vaccinations. A total of 185 participants responded to the questionnaire (response rate : 68%), and the period between receiving the initial vaccination and their response to the questionnaire ranged from 1.0 to 7.5 years (median : 4.0 years). The prevalence of breakthrough varicella after the initial vaccination was 17% among seroconverters who did not receive booster vaccination and 14% among non-seroconverters who received booster vaccination, showing no significant difference between the two groups. In conclusion, there are no safety issues regarding the administration of a booster vaccination to children with PVF after an initial varicella vaccination, and,a good antibody response can be expected.

Nosocomial transmission of varicella to a healthcare provider positive for anti-varicella zoster virus antibodies: nonprotective positivity with an immune adherence hemagglutination assay

A 24-year-old male healthcare provider, having attended a varicella patient 2weeks before, developed varicella himself. He had shown a positive result for anti-VZV antibodies measured with an immune adherence hemagglutination assay (1:4) 1year before. The present case shows that a positive result with this assay does not necessarily indicate protection against VZV infection.

Relative frequency of rotavirus subgroups 1 and 2 in Venezuelan children with gastroenteritis as assayed with monoclonal antibodies.

Monoclonal antibodies recently developed against the 42,000-dalton protein of two rotavirus strains were used in an enzyme-linked immunosorbent assay to determine the subgroup specificity of 252 specimens collected during a 45-month period from Venezuelan children with rotavirus gastroenteritis. Subgroup 2 rotavirus was shed by 85% of the children, whereas only 14% shed subgroup 1 rotavirus (one-half of them in a 3-month period). No differences were found in the occurrence of fever and vomiting between children shedding either rotavirus subgroup, but it appeared that the syndrome tended to last longer in children shedding subgroup 2 rotavirus. The monoclonal subgrouping enzyme-linked immunosorbent assay seemed to be more sensitive than an immune adherence hemagglutination assay, an enzyme-linked immunosorbent assay with polyclonal antibodies, or the electrophoretic analysis of RNA extracted from the virus. Overall, 99% of the specimens could be subgrouped by this assay.

Tumor-associated antigen from a human breast tumor cell line

Membrane-bound antigens were solubilized and purified from the continuously growing human mammary adenocarinoma cell line SW527. Two distinct breast tumor-associated antigens (BTA-1 and BTA-2) were obtained, as well as purified HLA antigens. The molecular weights of BTA-1 and BTA-2 were estimated at 112,000 and 65,000, respectively, by analytical polyacrylamide gel electrophoresis. The two mitogens also differed in carbohydrate content and hexose/pentose ratio. Antibodies to these antigens, raised in rabbits, were shown to be directed against SW527 surface markers by an immune adherence hemagglutination assay. Purified BTA-1 and BTA-2 reacted with rabbit antibodies to SW527 and with antibodies to a tumor-associated antigen previously isolated from tumor tissue freshly obtained from a patient with mammary adenocarcinoma, but did not react with anti- β 2-microglobulin. By indirect immunofluorescence assay, SW527 surface markers were shown to be recognized by antibodies in the serum of a patient with mammary adenocarcinoma; this reactivity was blocked by treatment of the serum with the tumor-associated antigen purified from fresh tissue. These results indicate that the antigens isolated from tumor cells in tissue culture and directly from human tumor tissue have common immunological properties.

EBV-IgA and new heterophile antibody tests in diagnosis of infectious mononucleosis.

An evaluation has been made of the EBV-IgA tests and the immune adherence hemagglutination assay (IAHA) as compared with standard diagnostic procedures in 119 serial sera from 22 clinical and 113 sera from 42 subclinical cases of EBV infectious mononucleosis. EBV-IgA antibody was demonstrable in 86.4 per cent of patients using EB3 cells as antigen and in 68.2 per cent with P3/HRIK cells. For heterophile antibody the IAHA test was more sensitive, gave higher titers, and was positive longer than the standard absorbed horse or sheep RBC tests in both clinical and subclinical EBV infections.

Detection of antibodies to Epstein-Barr virus capsid antigen by immune adherence hemagglutination.

The immune adherence hemagglutination assay was found to be as sensitive and specific as the indirect immunofluorescence technique for titration of antibodies to Epstein-Barr virus capsid antigen. Satisfactory virus capsid antigen-specific and negative control antigens for the immune adherence hemagglutination assay were prepared from cell extracts of the Epstein-Barr virus producer P3HR-1 and the Epstein-Barr virus genome-negative BJAB lymphoblastoid cell lines, respectively. As the immune adherence hemagglutination assay can be used to titrate antibodies to both the heterophil antigen of the Paul-Bunnell type and to virus capsid antigen, it offers a promising alternative to the immunofluorescence methods in the serodiagnosis of Epstein-Barr virus infections which can be performed by most diagnostic laboratories.

Antigenic characterization of human and animal rotaviruses by immune adherence hemagglutination assay (IAHA): evidence for distinctness of IAHA and neutralization antigens.

An immune adherence hemagglutination assay (IAHA) and a modified enzyme-linked immunosorbent assay for antigenic characterization of human rotaviruses were developed. The designations of type 1 and type 2 were identical to those established previously by specific complement fixation, enzyme-linked immunosorbent assay, and immune electron microscopy. By IAHA (and modified enzyme-linked immunosorbent assay) certain animal rotaviruses were found to be closely related to human rotavirus type 1. The pattern of IAHA reactivity and the cell culture neutralization serotype were found to be distinct properties. The separation of neutralization and IAHA reactivity was apparent when animal rotaviruses which were distinguishable from each other by neutralization assays were found to share IAHA specificity. Further evidence for the dissociation of the neutralization and IAHA specificities was found in studies of human and bovine rotaviruses which underwent genetic reassortment during coinfection. Thus, it appeared that the IAHA and neutralization antigens were coded for by different genes. In view of these findings, we suggest that the term serotype be reversed to identify the antigen that reacts with neutralizing antibodies as is customary for other viruses and that the term subgroup (instead of serotype) be used for the specificity detected by specific complement fixation, enzyme-linked immunosorbent assay, and now IAHA.

Genes of human (strain Wa) and bovine (strain UK) rotaviruses that code for neutralization and subgroup antigens

Sixteen rotaviral reassortants recovered from mixed infection with a ts mutant of bovine rotavirus (UK strain) and noncultivatable or cultivatable human rotavirus (Wa strain) were genotyped by polyacrylamide gel electrophoresis. Analysis of the reassortants showed that the ninth RNA gene segment regularly segregated with the neutralization specificity and the sixth RNA gene segment regularly segregated with the subgroup antigenic specificity detected by immune adherence hemagglutination assay. This indicates that the ninth RNA segment codes for the protein that induces and reacts with neutralizing antibodies while the sixth RNA segment codes for the subgroup antigen. In addition, it appears that the fourth and/or fifth RNA segments may be responsible for restriction of growth of the noncultivatable human rotavirus in cell culture.

Serum Antibody Levels as Risk Factors in the Dissemination of Herpes Zoster

Serum antibody levels against varicella-zoster virus (VZV) were examined by immune adherence hemagglutination assay (IAHA), indirect fluorescent antibody (IFA) assay, and complement fixation techniques in 67 immunocompromised patients with localized and disseminated herpes zoster. In the serum obtained initially, undetectable IAHA titers were found in 56.5% of the patients with disseminated zoster compared with 18.2% of those with localized zoster. When serum obtained within the first seven days of illness was analyzed, undetectable IAHA titers and IFA titers of less than 32 were noted in 77.8% of those with disseminated zoster but in only 18.5% of those with localized disease. Peak serum antibody titers in patients with disseminated zoster were eventually equal to or greater than those in localized zoster. The patient groups were comparable in age, underlying disease, and therapy, although Hodgkin's disease was more frequent in patients with disseminated zoster. Thus, the absent IAHA or low IFA levels of circulating antibody early in illness were highly significant risk factors in dissemination of virus in herpes zoster.

Antibody to early antigens of varicella-zoster virus during varicella and zoster.

IgG antibody to the early antigens of varicella-zoster virus (VZV) was studied during both varicella and zoster by the indirect immunoperoxidase antibody technique. In parallel, complement-fixing, immune-adherence hemagglutinating, IgG, and IgM antibodies to VZV were studied. In both varicella and zoster infections, antibody to the early antigens of VZV appeared three to five days after onset of infection, reached a peak during the second week, and progressively decreased in titer until it disappeared, usually within two months. This antibody usually appeared slightly later than IgG or IgM antibody and grossly correlated with IgM antibody in varicella. In zoster infections, IgM antibody to VZV was not found by the immune-adherence hemagglutination assay at a titer of greater than or equal to 1:4, whereas antibody to the early antigens showed a curve similar to that found in varicella. It is suggested that antibody to the early antigens of VZV be considered as a marker of acute VZV infection, which is associated with a specific and significant IgM antibody response in varicella but not in zoster.

Serologic response in human hepatitis A: detection of antibody by radioimmunoassay and immune adherence hemagglutination.

An indirect solid-phase radioimmunoassay (RIA) for detection of antibody to the hepatitis A antigen (anti-HAV) was developed using polystyrene pearls as the solid phase and hepatitis A antigen (HAAg) extracted from marmoset livers. This RIA was compared to an immune adherence hemagglutination assay (IAHA) which employed HAAg derived from the stools of chimpanzees collected during acute hepatitis A. Anti-HAV was detected in the sera of 15 humans with naturally acquired hepatitis A infection. Sensitivity and specificity were greater using the RIA, permitting the detection of anti-HAV as early as the time of onset of jaundice. Either seroconversion or a significant increase in the titer of anti-HAV was demonstrated following hepatitis A exposure in paired sera from six patients by both techniques. No significant difference in anti-HAV responses was noted between patients with icteric compared to anicteric hepatitis A or between children and adults with hepatitis A.

Prevalence of antibody to the Norwalk agent by a newly developed immune adherence hemagglutination assay

An immune adherence hemagglutination assay (IAHA) for the detection of antibody to the Norwalk agent of acute epidemic nonbacterial gastroenteritis was developed using as antigen virus purified from stool from an experimentally infected volunteer. The assay was sensitive and specific and was efficient for detecting Norwalk antibody seroresponses. The prevalence of Norwalk antibody in various groups in the United States was studied. Antibody to the Norwalk agent was acquired gradually, beginning slowly in childhood and accelerating in the adult period so that by the fifth decade 50% possessed antibody. This pattern of antibody acquisition contrasted sharply with that for the human rotavirus of infantile gastroenteritis. Rotavirus antibody was acquired during early childhood by almost all individuals in the pediatric groups studied. Antibody to the Norwalk agent was also found in rural Bangladesh; in a small prevalence survey of 39 children and adults 21% possessed Norwalk IAHA antibody, whereas 95% possessed antibody to the human rotavirus.

Non-A, Non-B Hepatitis

Editorials1 July 1977Non-A, Non-B HepatitisBARUCH S. BLUMBERG, M.D.BARUCH S. BLUMBERG, M.D.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-87-1-111 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptThe term "non-A, non-B hepatitis" is applied to cases of what appears to be viral hepatitis that cannot be identified as hepatitis A, hepatitis B or with the hepatitis associated with Epstein Barr virus and cytomegalovirus. A series of papers describing this variety of hepatitis (1-5), including several recently published clinical and epidemiologic studies (6-10), have helped in the understanding of the disease. Antibody to hepatitis A virus can be detected by an immune adherence hemagglutination assay using hepatitis A antigen purified from human stool. The presence of antibody implies previous infection. A rise in anti-HA titer in a serum...

Detection of Cytomegalovirus Antibody by Immune Adherence Hemagglutination

SummaryThe principle of complement-mediated immune adherence was applied to development of an immune adherence he-magglutination (IAHA) assay for antibody to cytomegalovirus (anti-CMV). Compared to complement fixation (CF), IAHA was two to eight times more sensitive for the detection of anti-CMV. In paired serum samples from multiply transfused patients, the concordance between rises in anti-CMV titers detected by CF and IAHA was 0.671 (P < 0.001). Convalescent titers measured by IAHA were significantly higher than those determined by CF in serum pairs in which anti-CMV rises were demonstrated by both techniques, and IAHA more commonly revealed anti-CMV in pretransfusion serum samples. No difference was noted between the kinetics of CF and IAHA antibody development after transfusion. Immune adherence hemagglutination, we conclude, is an efficient, sensitive alternative to CF for the detection of anti-CMV.The authors thank Mrs. Virginia L. Frye for preparing the typescript, and Dr. David W. Ailing for assi...

Hepatitis B surface antigen in the serum of infants after delivery from asymptomatic carrier mothers

A survey of sera of 5,993 pregnant women for hepatitis B surface antigen in Tokyo revealed 139 asymptomatic carriers (2.3%), essentially the same as that of a control population of comparable age group (2.2%). None of 59 specimens of cord blood of their newborn infants was positive for HBsAg according to an immune adherence hemagglutination assay. In 11 mother-child pairs in whom follow-up was possible for more than seven months, HBsAg was detected in the sera of eight infants within the first six months, after birth, with antigenemia persisting throughout the observation period, while antigenemia was not detected in the remaining three. The subtype of HBsAg was identical for each mother-child pair. The antigen was detected in sera of two of the infants after appropriate incubation periods of 123 and 133 days, respectively, whereas in others it was detected as early as 5 and 13 days after delivery.