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PubMed1979

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.

Adult↗
PubMed1990

[Phagocytic capacity of peritoneal exudate cells from rats immunized with a ribosomal preparation of Ty2 Salmonella typhi].

It was compared the activity of exudate peritoneal cells (EPC) obtained from CFW mice immunized either with Salmonella typhi Ty2 ribosomal fraction or whole-cell heat inactivated vaccine, both in comparison with EPC from sham-immunized. In the group which received ribosomal preparation, a subcutaneous dose equivalent to 100 micrograms of RNA in incomplete Freund's adjuvant (IFA) was initially used and 14 days after a booster of the same dose in IFA was given. A single dose of whole-cell heat inactivated vaccine, with 10(6) bacteria in IFA was employed subcutaneously in animals of the second group. EPC from controls and immunized mice were withdrawn at periods of 7, 11, 14, 18, 22, 25, 29 and 31 days after immunization and each sample was incubated in vitro in presence of live virulent non-opsonized S. typhi Ty2 in 1:200 cell-bacteria relation. Twenty four hours after cultivation, EPC bacterial capacity was determined after cell disruption and enumeration of survival bacteria were made through viable counts. Results have shown that EPC from mice immunized were more efficient in eliminating intracellular bacteria than those which came from sham-immunized animals. Also, it was found that EPC from mice immunized with ribosomal preparation were more efficient (maximum P = 0.005) than EPC from the mice which received killed whole bacteria.

Animals↗
PubMed1989

[False positive results of HIV virus tests in patients undergoing chronic hemodialysis].

The sera of 173 haemodialysis patients treated in two dialysis centers in Hungary were tested for the presence of HIV (HTLV III/LAV) antibodies. Four different commercial enzyme immunoassay (EIA) kits and two types (CEM/LAV, and H9/HTLV III) of indirect immunofluorescence assay (IFA) were used. The Western blot technique was applied as confirmatory test in the study. No confirmed positive results were found in any of the cases. However, in 15 patients (8.7%) false positive (not confirmable by the Western blot assay) results were obtained in at least one but mostly in all of the three type 1 EIA kits (ORGANON, ELECTRONUCLEONICS, SORIN) applied. In 4 patients, the IFA assay also gave false positive results which could be repeated in sequential samples taken from the same patients. Increased reactivity in the control plate (coated with a concentrate of cellular material shed by uninfected H9 cell line) of the SORIN kit was found only in a few false positive samples and no fluorescence with the uninfected H9 or CEM cells was observed in any of the sera showing a false positive IFA. These results indicate that the false positive anti-HIV results frequently observable in haemodialysis patients are not simply the consequence of the presence of antibodies reacting with the uninfected H9 and/or CEM cells but they are most probably due to antibodies against antigens expressed on these cells only after infection with the human immunodeficiency virus.

False Positive Reactions↗
PubMed1989

[Immunological humoral response in mice experimentally infected with Echinococcus granulosus, preliminary result of kinetic observation].

5 serological methods included latex agglutination test (LA), enzyme-linked immunoelectrotransfer blot (EITB), enzyme-linked immunosorbent assay (ELISA), dot-enzyme-linked immunosorbent assay (Dot-ELISA) and immunofluorescence antibody test (IFA) were used to trace the kinetic changes of the humoral response in NIH mice experimentally infected with protoscolex of Echinococcus granulosus. Results showed that Dot-ELISA and IFA were able to detect antibodies 2 wk after infection while ELISA, EITB and LA gave positive reactions 4 wk, 2 mos and 3 mos after infection respectively. Therefore Dot-ELISA and IFA were considered to be the most sensitive ones among the 5 methods used. All infected mice gave positive reactions at 8 and 12 mos but none of the normal control mice showed any reaction throughout the period of observation (Fig. 1).

Animals↗
PubMed1989

Comparative analysis of ELISAs employing repetitive peptides to detect antibodies to Plasmodium falciparum sporozoites.

In the last few years, a number of different recombinant and synthetic peptides consisting of the repetitive sequence of the Plasmodium falciparum circumsporozoite protein (NANP)n have been produced and used to develop immunoassays for the detection of antibodies against P. falciparum sporozoites in human sera. A comparative study of three enzyme-linked immunosorbent assays (ELISAs) that employed different (NANP)n peptides (the synthetic peptides (NANP)3 and (NANP)40 as well as the recombinant peptides R32tet32 and R32LR) was carried out using serum samples from individuals who were living in different malaria-endemic areas. The results obtained for these peptide-based ELISAs were compared with those obtained for an immunofluorescence assay (IFA) that used glutaraldehyde-fixed sporozoites. All the methods tested exhibited 100% specificity on sera from persons not exposed to malaria, good reproducibility (coefficients of variation ranged from 3% to 15% for peptide-based ELISAs), and good sensitivity. Reproducibility and sensitivity were lower for the IFA than for the peptide-based ELISAs, perhaps because of the subjective element in the interpretation of the results which is inherent in the IFA method. ELISAs based on peptides that contain a higher number of (NANP) repeats, i.e., (NANP)40 and R32tet32 or R32LR, gave results which correlated better with each other than with those obtained with the ELISA that employed a shorter (NANP)3 peptide. (NANP)n-based ELISAs are relatively simple and inexpensive methods for the detection of anti-P. falciparum sporozoite antibodies and can readily be used in epidemiological research in the field. These assays could contribute to a better understanding of the natural history of the host-parasite relationship in malaria research.

Animals↗
PubMed1989

Identification of Tritrichomonas foetus in sections of bovine placental tissue with monoclonal antibodies.

Sections of bovine placenta from cases of bovine trichomoniasis were examined for the presence of Tritrichomonas foetus by standard histological methods using phase-contrast microscopy and by indirect immunofluorescence assay (IFA) employing monoclonal antibodies (mAbs) specific for T. foetus. Parasites were identified readily in deparaffinized tissue up to 4 yr old by IFA with 2 mAbs previously shown to bind to the surface of living T. foetus. These results indicated that the IFA provided a rapid and specific method of identifying T. foetus in tissue sections as compared to standard histological methods.

Abortion, Veterinary↗
PubMed1989

Assessment of putative tests for protective immunity to falciparum malaria.

Sera from clinically immune individuals comprising 10 hospitalised patients (Group I), 30 persons residing in a malaria endemic area in Thailand (Group II) and 8 persons from a hyperendemic area in Ivory Coast (Group III) were tested by the parasite growth inhibition (PGI), indirect fluorescent antibody test of ring-infected erythrocyte surface antigen (RESA-IFA), urease-ELISA and Western blot. Paired sera from patients recovering from malaria (Group IV) as well as sera from blood donors were also tested. In the PGI test, sera were tested against three uncloned isolates of P. falciparum comprising SO, I4 and AE9 (PGI-SO, PGI-I4 and PGI-AE9 respectively). When growth inhibition of greater than or equal to 30% against any one of the three isolates was considered positive, the positive rate for the combined Groups I, II and III was 78.7%. Further analysis showed that the positive rates for PGI-SO, PGI-I4 and PGI-AE9 were 63.8%, 59.5% and 59.5% respectively and were not significantly different (p greater than 0.05). Comparison between PGI-SO, PGI-I4 and PGI-AE9 activities of Groups I, II and III sera showed no significant differences in any comparison groups except with PGI-AE9 in which Group III sera were more frequently positive than Group II sera (p = 0.004). Follow-up of PGI-SO and PGI-AE9 activities in Group IV patients showed mostly a decrease or no change in the activities of the convalescent sera taken 63 days later. RESA-IFA positive rate in the combined Groups I, II and III sera was 91.7%. There were no significant differences either in the seropositive rates or in the geometric mean antibody titers (GMT) between Groups I, II and III sera. Follow-up in Group IV patients showed no change in antibody titers in 64% of cases, decrease and increase in titers in 29% and 7% of cases respectively. The urease-ELISA seropositive rate in the combined Groups (I, II and III) was 89.5% which is not significantly different from that of RESA-IFA (p greater than 0.05). Comparison between individual Groups (I, II and III) likewise showed no significant differences in both GMT and seropositive rates. Follow-up in Group IV sera showed either no change or a decrease in antibody titers in 55.6% and 44.4% of cases respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗
PubMed1989

A comparison of the ELISA with other serological tests in the serodiagnosis of amoebiasis.

The sera of 78 patients with invasive amoebiasis were tested for antiamoebic antibodies by the techniques of enzyme-linked immunosorbent assay (ELISA), indirect haemagglutination (IHA), indirect fluorescent antibody (IFA) and counterimmunoelectrophoresis (CIEP). Results showed that the ELISA compared favourably with IHA and IFA tests in terms of sensitivity and specificity. ELISA, IHA and IFA detected 97.4%, 96.2% and 98.7% of the patients respectively. CIEP was the least sensitive of the 4 serological methods with a sensitivity of 88.5%. The advantages and disadvantages of the 4 serodiagnostic procedures are discussed.

Amebiasis↗
PubMed1989

[Antimalarial serology in 1987-1988 in the Highland Plateaux, the East coast and the South of Madagascar].

We studied 521 serum samples collected in 1987-88 in the Highland Plateaux and the East coast of Madagascar, two areas presenting different levels of malaria endemicity. Total anti-Plasmodium falciparum antibodies were investigated by an indirect immunofluorescence assay (IFA). Antibodies directed to the ring-infected erythrocyte surface antigen (RESA) were investigated by a modified IFA (MIFA). Results were analysed in regards to malariological parameters (parasite and splenomegaly rates) collected simultaneously. IFA appears as a good epidemiological tool as it closely parallels the classical malariological parameters. In selected studies, the presence of P. falciparum parasites in blood was less frequent in individuals presenting with anti-RESA antibody, as detected by MIFA, than in the other individuals were consistently lower in subjects with anti-RESA antibody than in others.

Animals↗
PubMed1985

Serologic evaluation of vaccinated American river otters.

The Oklahoma Department of Wildlife Conservation acquired 20 American river otters (Lutra canadensis) between 1984 and 1985 for reintroduction into Oklahoma waterways. In 1985, 10 otters were evaluated for serum antibody titers after vaccination with canine distemper virus, canine adenovirus type 2, canine parvovirus (CPV), feline panleukopenia virus (FPV), feline rhinotracheitis virus (FRV), and feline calicivirus. Prevaccination serum-virus neutralization (SVN) antibody to feline rhinotracheitis virus was found in 2 otters and to feline calicivirus in 1 otter. Using an indirect fluorescent antibody (IFA) assay, prevaccination antibody to CPV and FPV was found in 2 otters. A significant increase in SVN antibody titers was found after vaccination of otters with canine adenovirus type 2 (6 of 8 animals) and feline calicivirus (1 of 8 animals). One of 8 otters developed significant antibody titers to CPV and FPV, as measured by IFA assay. Otters did not develop SVN antibody titers to canine distemper virus after vaccination. Antigens of feline leukemia virus, using ELISA, or antibodies to feline infectious peritonitis, using IFA assay, were not found in the 20 otters.

Adenoviridae↗
PubMed1986

Canine parvovirus: development of immunofluorescence and immunoperoxidase techniques.

Two methods of immunocytochemistry, immunofluorescence (IFA) and immunoperoxidase (PAP) were used to demonstrate canine parvovirus (CPV) antigens in sections of canine tissue. Specific staining using IFA and PAP was successful only in sections of fresh frozen tissue and formalin fixed/formol sublimate postfixed tissues respectively. A range of tissues was then taken at post mortem examination from a puppy which had been experimentally infected with CPV. Upon comparison, PAP staining gave high resolution, a permanent preparation and clear intracellular localisation of antigen. IFA resulted in less defined localisation of antigen but the technique was simpler and more easily controlled.

Animals↗
PubMed1987

[Feline leukemia virus (FeLV) and FeLV-associated diseases in cats: a review].

Feline leukaemia virus (FeLV) usually occurs in its natural species, the domestic cat. FeLV is also important to human individuals as a comparative model, as FeLV may cause a variety of diseases which are partly malignant and partly benign, such as immunosuppression which bears a resemblance to AIDS (acquired immune deficiency syndrome) in man. Although FeLV is a common infective agent, the incidence of disease due to FeLV is much higher in cats kept in closed households in which several of them are present than it is in free-range cats. Consequently, diseases caused by FeLV are frequently diagnosed in pedigree cats which are often maintained in relatively large numbers. FeLV is transmitted among cats by contagion. The main sources of infection are persistantly infected FeLV carrier cats which continuously excrete virus from the mouth and in other secretions. The majority of cats infected with FeLV will produce neutralising antibodies. Cats which are unable to do so, will become permanently infected. The prognosis is bad in these cats: 90 per cent will die within five years. Various techniques are used to detect FeLV. The most common method, the indirect immunofluorescence (IFA) test, is performed on air-dried blood smears. The results of the IFA agree with that are almost completely identical to those of the virus isolation test. Another test is ELISA (enzyme-linked immunosorbent assay), which produces approximately 10 per cent more positive results which are probably due to circulating FeLV antigens. Dissemination of FeLV among cats may be prevented by identifying infected carrier cats and removing them from contact with non-infected cats. Removal programmes using indirect immunofluorescent antibody (IFA) tests were used successfully in the Netherlands. The proportion of FeLV-positive cats decreased from 9 per cent in 1974 to approximately 3 per cent in 1985 during such a removal programme. During the above period, the removal programme was carried out in the society of Dutch cat breeders 'Felikat', the programme being made compulsory on all members of the society. The incidence of cats positive for FeLV decreased from over 11 per cent in 1974 to less than 2 per cent within four years. None of the cats tested in this society was found to be positive for FeLV in 1984 and 1985. Besides removal programmes, other methods of control, such as vaccination, were developed to prevent the spread of FeLV. The FeLV-immunostimulating complex vaccine (FeLV-ISCOM vaccine) a subunit vaccine in which FeLV-gp70 is presented in a particular manner, seems to be promising.

Animals↗
PubMed1988

Comparison of haemagglutination test, enzyme-linked immunosorbent assay and indirect immunofluorescence antibody test for determination of rubella immune status.

Comparative evaluations of immune status for rubella are described for the enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence (IFA) and two standard haemagglutination inhibition tests (HAI kaolin; HAI heparin-MnCl2). In general, a reasonably good correlation was obtained between the level of rubella antibodies measured by the HAI (kaolin) test and by ELISA, but an appreciable proportion (15%) of ELISA positive specimens were encountered among HAI (kaolin) negative sera. All of these HAI negative, ELISA positive sera, except one were found to be positive for rubella antibodies by IFA. Neutralization test performed on this serum positive by ELISA only, confirmed the presence of protective rubella antibodies. Of all the tests evaluated ELISA appeared unequivocally to be the most sensitive test followed closely by IFA. The standard HAI (heparin-MnCl2) was more suitable than the HAI (kaolin), for the determination of immune status. Further, no linear relationship between single rubella virus HAI and ELISA values was observed.

Antibodies, Viral↗
PubMed1988

Identification of a surface antigen on Loa loa microfilariae the recognition of which correlates with the amicrofilaremic state in man.

Filarial infections induce a spectrum of disease in their natural hosts, and by correlating immunity found in individuals with their disease pattern, one may delineate non-pathogenic, protective mechanisms. Loa loa is causal of mild to moderate pathology, and it is unique among the human filaria in that adult worms are occasionally visible during subconjunctival migration. To study immune mechanisms controlling microfilaremia, sera from 15 subjects with amicrofilaremic occult loiasis (OL) were compared with sera from 10 subjects with microfilaremic loiasis (ML) microfilaremia, (greater than 4000/ml) for their reactions with living microfilariae (mf). An IFA was first used to detect antibodies able to bind to the surface of living L. loa mf. ML subjects either did not react (7/10) or reacted only very weakly (3/10). Highly reactive sera were found only in OL subjects; 7/15 gave very bright fluorescence, 5/15 gave moderate reactions, and 3/15 were negative. Most of these antibodies were of the IgG class. Sera from all subjects were also reacted with living mf in an antibody-dependent cellular adherence test using normal leukocytes. Sera that were strongly positive in IFA showed strong adherence and IFA-negative sera were non-reactive. To identify the Ag involved, mf were surface iodinated, detergent-extracted Ag were immunoprecipitated, and Mr was determined on SDS-PAGE. Several OL sera, all highly reactive in the above tests, precipitated a 23-kDa molecule with which all ML sea failed to react. Sera from a mandrill experimentally infected with L. loa also precipitated the 23-kDa Ag when taken post-patency. In conclusion, it appears that certain people who control L. loa microfilaremia have high levels of IgG antibodies that bind to a surface Ag of 23 kDa and are able to mediate cellular adherence.

Adhesiveness↗
PubMed1986

Serological cross-reactions between Rickettsia typhi, Proteus vulgaris OX19, and Legionella bozemanii in a series of febrile patients.

The sera of 19 patients with a febrile disease of undetermined etiology were positive in the indirect immunofluorescence assay (IFA) to Legionella bozemanii serogroup 1 (Lb) and Rickettsia typhi (Rt). To both antigens, high titers of IgG-class and IgM-class antibodies were demonstrated. Several of the patients also had positive IFA and Weil-Felix reactions to Proteus vulgaris OX19 (PX 19). A sharp reduction of the serotiters to all three antigens was achieved by absorption of the sera with any one of the organisms. We demonstrated, by crossed immunoelectrophoresis with an Lb extract and a rabbit reference anti-Lb serum, that a heat-stable and trypsin-resistant antigen (antigen no. 1) reacted consistently with patients' sera that had been incorporated into an intermediate gel. Sera from five patients with high-titer IFA reactions to Rt, but no reaction to Lb, showed no interaction with antigen no. 1.

Antigens, Bacterial↗
PubMed1977

Antivascular endothelial cell antibody--its role in transplantation.

Sera samples from eight different groups of patients were tested for the presence of circulating antibody (IgG) directed against vascular endothelial cell antigens. The indirect immunofluorescent method which used either whole blood vessels or single endothelial cells as targets was compared to lymphocytotoxicity panels. The indirect immunofluorescent antibody test (IFA) consistently detected antibody in sera samples which were negative for lymphocytotoxic activity, and the presence of IFA antibody to vascular endothelial cells had a much better correlation with both clinical course and renal allograft rejection than did the lymphocytotoxicity panels. Single vascular endothelial cells appeared to be a more sensitive target for the detection of IFA antibody than did whole vessels. Preliminary absorption studies suggest that cell-specific as well as HL-A antigens play a role in the immunological response to vascular endothelial cells.

Antibodies↗
PubMed1988

[Diagnostic bronchoalveolar lavage in assessing the efficacy of the treatment of idiopathic fibrosing alveolitis].

A study of the results of repeated diagnostic bronchoalveolar lavage in 27 patients with idiopathic fibrosing alveolitis (IFA) has shown that the investigation of cellular composition of the bronchoalveolar content (BAC) made it possible to specify IFA activity and its therapy-related change. Normalization or a decrease below 6% of the neutrophil content in BAC was typical of achieving low activity and remission of disease. A relatively high count of neutrophils in BAC (over 10%) during therapy was observed in preserved significant IFA activity resistant to drug therapy.

Adult↗
PubMed1977

The serological relationship of a British Theileria with other Theileria species using the indirect fluorescent antibody test.

Antisera and antigens of a Theileria species isolated from British cattle were compared with those of six strains of Theileria mutans from geographically separated areas in East and South Africa, using the indirect fluorescent antibody (IFA) test. It was found that the British Theileria species did not react in the IFA test with these strains of T mutans. The British Theileria species was also compared with a variety of other Theileria species using the IFA test and no reactions were detected. It was concluded that the British Theileria species, which has been designated T mutans Essex, could be distinct from African T mutans.

Animals↗