Rapid immunofluorescence diagnosis of respiratory syncytial virus infections among children in European countries.
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Child, Preschool↗
Biomedical subjects
M Grandien
Publications and source records attributed to M Grandien.
Rubella immunity testing by mixed hemadsorption.
A mixed hemadsorption (MH) test for naturally acquired rubella immunity and immune responses after rubella vaccination has been compared with the hemagglutination-inhibition (HI) test, the immunodiffusion (ID) test, the neutralization test (NT), the hemolysis-in-gel (HIG) test and the indirect immunofluorescence (IF) test. The MH test was approximately equivalent to the HI test and the NT as to sensitivity for the early response to rubella vaccination, and all three tests are thought to measure protective antibodies, i.e. antibodies supposedly directed against envelope antigens, to about the same extent. The MH test for rubella antibodies is especially suited for laboratories in which this type of test is routinely used for other purposes, e.g. rabies, respiratory syncytial virus, herpes simplex, varicella and certain auto-antibodies. Where the method is already standard it may be preferable to the NT which, although of similar clinical value requires tedious standardization. Both tests may be necessary in cases where the HI test is equivocal or hampered by non-removable on-specific inhibitors.
Antibody Formation↗
Preparation of sera for subtyping of influenza A viruses by immunofluorescence.
The conditions for preparation of type-specific and subtype-specific influenza A virus reagents to be used in the immunofluorescence technique have been evaluated. Type A-specific antibodies were prepared by passing an antivirion hyperimmune serum through an immunoadsorbent column containing antigens from disrupted virions of a different influenza A virus subtype. The type-specific antibodies were recovered from the immunoadsorbent by desorption with 3 M NaI. For subtype determination, antisera against the various hemagglutinins were used. Such sera could be prepared by removal of irrelevant influenza A virus antibodies from sera directed against purified virions and isolated peplomers, respectively. This was performed by passing the antisera through immunoadsorbent columns containing antigens from disrupted virions of appropriate strains. However, attempts to obtain a hemagglutin-specific antiserum from a serum directed against allantoic fluid virus suspension failed with this procedure. Antisera obtained after immunization with purified hemagglutinin were also elaborated. These sera were shown to be superior for subtyping of influenza A virus infections by immunofluorescence, but could not a priori be regarded as subtype-specific. The usefulness of subtype-specific sera has been demonstrated on clinical specimens for rapid virus diagnosis.
Animals↗
Progressive rubella virus panencephalitis: synthesis of oligoclonal virus-specific IgG antibodies and homogeneous free light chains in the central nervous system.
The occurrence of oligoclonal IgG and homogeneous free lambda light chains in the cerebrospinal fluid (CSF) of a patient with progressive rubella virus panencephalitis is reported. Fractions of oligoclonal IgG corresponding to those of the CSF occurred also in the serum. Evidence is presented that the oligoclonal IgG of serum and CSF represent rubella virus-specific antibodies. Different populations of oligoclonal IgG appear to be associated with antibody specificities to different antigenic components of the virus.
Adolescent↗
Early diagnosis of tick-borne encephalitis (TBE) by demonstration of specific IgM antibodies.
The demonstration of specific IgM antibodies after separation of immunoglobulins was used for the early diagnosis of tick-borne encephalitis (TBE). The separation was done by sucrose-gradient density ultracentrifugation and IgM antibodies were demonstrated with the hemagglutination-inhibition (HI) test. IgM antibodies were found in 9/11 sera taken within 3 days after onset of clinical symptoms and in all sera (37/37) taken 4--15 days after onset of the disease. Most sera (22/24) were still IgM positive 3--6 weeks after onset of the disease. After this period of time IgM antibodies could no longer be defected. The demonstration of IgM antibodies also proved valuable in confirming the diagnosis in several cases where no antibody rise was found neither by the complement-fixation (cf), nor by the HI test.
Antibodies, Viral↗
Serum Q fever antibodies in Swedish UN soldiers in Cyprus--reflecting a domestic or foreign disease?
Swedish soldiers from 2 battalions serving in Cyprus 1975 and 1976 were tested serologically for Coxiella burnetii phase 2 complement-fixing antibodies. 2.7% were positive. Although none of the seropositive soldiers had been seriously ill abroad, the titers are considered to emanate from an infection in Cyprus as there is no evidence of domestic Q fever from a survey of 1390 Swedes.
Antibodies, Bacterial↗
Comparison of immunofluorescence and immunoperoxidase methods for viral diagnosis at a distance: a WHO collaborative study.
Nasopharyngeal secretions collected in Newcastle were examined in both Newcastle and Stockholm for the presence of influenza virus type A and respiratory syncytial (RS) virus by immunofluorescence and immunoperoxidase techniques. A total of 139 specimens were examined in this way and the agreement between the two centres for immunofluorescence was 94% for influenza virus A, 95% for respiratory syncytial virus, and 95% for negative specimens. This technique can therefore be used for examining specimens taken at great distances from the collecting laboratory. The results of the immunoperoxidase technique were unsatisfactory mainly because of the presence of endogenous peroxidase. Measures taken to remove this also destroyed RS virus antigen. It is premature to introduce the immunoperoxidase technique for rapid virus diagnosis, but in due course, when the problems outlined in this article are solved, it could become a useful technique.
England↗
The advantage and adequacy of the mixed hemadsorption test for rabies antibody determination.
The goal of rabies vaccination is an early development of protecting antibodies. The mixed hemadsorption (MH) test is shown to be an adequate test to evaluate the immunization response. It measures antibodies combining with antigens on rabies-infected cells equivalent to the glycoprotein antigen at the exterior of the budding virus (1). Further, the MH test used measures antibodies solely of IgG class, thus avoiding positive results being obtained by IgM antibodies which can be produced during a prolonged period because of antigen persistence due to daily immunization doses (2). Antibodies of IgM class may play only a minor role in the early defence against rabies as probably only antibodies of IgG class can reach a locally introduced virus and, so far, are the only immunoglobulins shown to induce immune lysis in rabies-infected cells (3). The MH test is described. It is a sensitive, reliable and technically convenient method as it has a capacity for testing many sera in the same run. Results obtained are compared with neutralization values obtained from different types of rabies vaccines (4).
Antibody Formation↗
Enhanced cytotoxicity of human lymphocytes against rabies-infected cells by rabies-specific antibodies.
Human anti-rabies immune sera enhanced the in vitro cytotoxicity of human lymphocytes against rabies virus-infected green-monkey kidney cells. The immune sera were collected from patients immunized with rabies vaccine produced either in human diploid cells or in nervous tissue. Significant cytotoxicity was observed even with high serum dilutions, indicating that the K-cell assay might be a sensitive tool for detection of anti-rabies antibodies.
Animals↗
Rapid diagnosis of influenza A infection by immunofluorescence. Methodological problems and clinical material.
Indirect immunofluorescence (IF) on cell spreads from nasopharyngeal secretions (NPS) was used for the rapid diagnosis of influenza A infection and was compared with IF on monkey kidney cells infected by NPS. The clinical diagnosis of influenza A infection was confirmed by serology in 32 of 40 patients. In 27 of the 32 patients (84 per cent), the diagnosis was achieved by IF on cell spreads of NPS. In 13 of 15 (87 per cent) subjects with positive serology, the early appearance of influenza A virus antigen was revealed by indirect IF on infected monolayer cells. No false positive specimens were found among serologically negative subjects by either method. Consequently, the reliability of IF on cell spreads of NPS is very similar to IF on infected cell cultures, but offers a much quicker diagnosis (3-4 h as compared to 1-3 d). Monovalent anti-human IgG FITC should be used instead of polyvalent anti-human Ig FITC, as the latter contains anti-IgA which may adhere to intracellular IgA in the epithelial cells of NPS and thereby cloak the viral antigen.
Adult↗
Evaluation of tests for rabies antibody and analysis of serum responses after administration of three different types of rabies vaccines.
Humoral antibody response to three types of rabies vaccines were assayed by the neutralization (NT), the mixed hemadsorption (MH), and the indirect immunofluorescence (IF) tests. The NT and MH tests were used to detect antibodies combining with antigens at the surface of virions and infected cells, whereas the indirect IF test measured antibodies mainly to the rabies nucleocapsid antigen. After immunization with a human diploid cell vaccine, antibodies were detected by both the NT and the MH test in the 14th- and 30th-day serum samples from each of eight vaccinated persons. There was a good correlation between titers obtained with the two tests in this group of vaccinees. Antibodies elicited by duck embryo and nervous tissue vaccines occurred less frequently and in lower titers. In these groups of vaccinees, 5 of 14 and 5 of 10, respectively, had antibodies detectable by the NT test in the 14th- and 30th-day sera but were negative by the MH test. It is suggested that this was due to the high levels of immunoglobulin M antibodies, which are known to be elicited by daily injections of vaccine. Since antibodies of the immunoglobulin M class are considered to be less important for protection against rabies, the MH test is recommended for immunity determinations. Compared with the NT test, this test also offers the advantage of being technically more convenient because of its capacity for testing numerous sera in a single run. Antibody titers obtained by the indirect IF test in the human diploid cell vaccine group were relatively low. Titers in the duck embryo and nervous tissue vaccine groups were higher but did not correlate with the results of the NT test.
Antibodies, Viral↗
New tests for characterization of mumps virus antibodies: hemolysis inhibition, single radial immunodiffusion with immobilized virions, and mixed hemadsorption.
Hemolysis inhibition (HLI), single radial immunodiffusion (SRID) with immobilized virions, and mixed hemadsorption tests were used for measuring antibodies against mumps virus. Rabbit hyperimmune sera against mumps and early and late human convalescent sera were analyzed. All three tests identified antibodies against both hemagglutinin and the second major envelope component, hemolysin (fusion factor). The sensitivity of the HLI test corresponded to that of the hemagglutination inhibition (HI) test, but in some sera HLI antibodies occurred in greater quantity than HI antibodies. The SRID test readily identified rises in antibody titers in connection with acute infection. Due to its simplicity and lack of sensitivity to nonspecific inhibitors, it is recommended for use in this context. The mixed hemadsorption test showed a high sensitivity for specific identification of mumps antibodies. It therefore may be suitable for use in screening for immunity to mumps.
Antibodies, Viral↗
Ornithosis as a nosocomial infection.
An outbreak of ornithosis in the department of infectious diseases of a general hospital is described. The outbreak comprised 12 cases aged 18-80 years. The index case had a history of contact with birds. He developed a serious illness and died. 11 persons contracted the disease after contact with the index case; 8 of them were personnel of the clinic of infectious diseases and 1 case was a patient hospitalized in the same room as the index case. All of the patients showed a typical pneumonia and one of them had symptoms of encephalitis. Treatment with doxycycline was successful. 200 healthy contacts were treated prophylactically with doxycycline. None of these displayed any symptoms of disease.
Adolescent↗
Antibodies to calf serum as a cause of unwanted reaction in immunofluorescence tests.
One potentially important cause of disturbing staining of cell cultures used for immunofluorescence (IF) tests was found to be a specific reaction between calf serum and corresponding antibodies. A simple method utilizing polymerized calf serum as an immunosorbent was used for the removal of the antibodies causing the undesired reactions. One to three cycles of absorption with the calf serum immunosorbent were highly efficient in removing antibodies to calf serum, whereas the titer of a heterologous antiserum (anti-egg albumin) was only slightly affected by the same procedure. An enzyme linked immunosorbent assay (ELISA) was used for determination of the efficacy of the absorption method.
Absorption↗
Evaluation of an immunodiffusion test for screening of rubella immunity.
Rubella antigen obtained by treatment with a mild neutral detergent (Brij-58) of rubella virus infected SIRC cells gave two lines of precipitate in a modified Ouchterlony test (ID), using sera collected during convalescence after rubella. One strong line was obtained with positive sera and sometimes one thin line as well. The presence of at least one line was taken as indicator of rubella immunity. The pattern in which reagents were arranged allowed all positive serum samples to give precipitates showing antibody identity with a rubella control precipitate. Among women in the fertile age screened for immunity, 75% gave precipitates by the ID test and thus were considered to be immune. This should be compared with the 85% positivity found by means of the haemagglutination-inhibition (HI) test. As the HI test occasionally gives false results due to non-removable, non-specific inhibitors, the ID test on basis of the specificity seems to be well suited to be used as a method for the selection of individuals to be vaccinated against rubella.
Adolescent↗
Determination of varicella immunity by the indirect immunofluorescence test in urgent clinical situations.
Certain categories of patients run an increased medical risk when exposed to varicella. Newborns lacking maternal immunity, immunosuppressed leukemic children or children with other immunodeficient disorders may gain a decisive benefit by administration of e.g. interferon or of specific immunoglobulin against varicella. An indirect immunofluorescence (IF) method has been used to estimate varicella. An indirect immunofluorescence (IF) method has been used to estimate varicella immunity. Of 51 heavily exposed women denying earlier varicella, 39 were found to be immune by the test and none of those contracted varicella, whereas 9 out of the 12 non-immune women fell ill. 14/48 immunosuppressed children were found to be seropositive and none of these contracted varicella, whereas 13 mild or subclinical cases occurred among the 34 seronegative children. The same test revealed that 175/182 (96%) young adults in Sweden were immune. The indirect IF test is considered a rapid, simple and reliable method for estimating varicella immunity.
Antibodies, Viral↗
Early diagnosis of virus-caused vesicular rashes by immunofluorescence on skin biopsies. II. Poxvirus (vaccinia).
Immunofluorescence (IF) was used to demonstrate vaccinia virus antigen in frozen sections of skin biopsies from the site of revaccination in 42 individuals. The immunoglobulin (Ig) of a rabbit anti-vaccinia serum and the Ig of the pre-immune serum conjugated with fluorescein-isothiocyanate (FITC) was employed. 11/13 biopsies taken 1 day after vaccination were positive in the IF test as were 13/13 biopsies taken 2 days and 14/16 biopsies taken 3 days after vaccination. Even minute quantities of virus antigen were easily detected. The applicability of the test and the advantage of using biopsy material in early rashes of vaccinia and variola is discussed. The reliability of the direct IF using conjugated antisera against vaccinia-variola, varicella-zoster and herpes simplex virus for differentiating between maculopapular rashes was proved in a coded test.
Antigens, Viral↗
Early diagnosis of virus-caused vesicular rashes by immunofluorescence on skin biopsies. I. Varicella, zoster and herpes simplex.
The use of immunofluorescence (IF) for the rapid identification of varicella-zoster (V-Z) and herpes simplex (HS) antigen in frozen sections of biopsies of early non-vesicular skin lesions was investigated. Direct IF, using fluorescein- isothiocyanate- conjugated immuoglobulin (FITC Ig) of paired human anti-V-Z sera and FITC-conjugated negative and positive anti-V-Z-monkey Ig, yielded specific fluorescence of virus antigen in all 14 varicella cases investigated and in 10/11 zoster cases. In contrast, indirect IF, using paired human anti-zoster sera and a sheep anti-human Ig FITC, was not satisfactorily specific, since staining with the anti-human Ig FITC alone also yielded fluorescence of infected cells in some cases. In 6/8 cases of HS infection, specific fluorescence of virus antigen was obtained by direct IF, using FITC-conjugated negative and positive guinea-pig anti-HS Ig. Because of the often predominant distribution of virus antigen to the corium and the skin appendages, punch biopsies are apparently better than scraped material, at least in the prevesicular stage.
Antigens, Viral↗