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PubMed1982

Avian reovirus antibody assay by indirect immunofluorescence using plastic microculture plates.

An indirect fluorescent antibody test was developed to detect serum antibody to avian reovirus strain WVU2937. This test employed small multiple well plastic plates (8 x 5.5 cm) which readily fitted into the standard mechanical stage mechanism of an incident light fluorescence microscope. The small wells of the plates required minimal (10 muL) volumes of reagents. In tests on 18 sera in which the indirect fluorescent antibody, agar gel precipitin and plaque reduction methods were compared sera which gave negative results in the agar gel precipitin test were sometimes positive in the indirect fluorescent antibody and plaque reduction test, but indirect fluorescent antibody titers were lower than plaque reduction test titers. No false positive reactions were detected in 46 sera from uninoculated specific pathogen free chicks of up to eight weeks of age.

Animals↗
PubMed1983

Serological diagnosis of Staphylococcus aureus septicaemia and endocarditis by means of crossed immuno-electrophoresis.

The diagnostic sensitivity and specificity were determined for serological diagnosis of Staphylococcus aureus endocarditis and septicaemia by means of crossed immuno-electrophoresis with intermediate gel. The antibody responses to S. aureus antigens in sera from 19 patients with S. aureus endocarditis and 51 patients with S. aureus septicaemia were compared with findings in 30 patients with non-S. aureus endocarditis and 30 patients with non-S. aureus septicaemia. Two of 55 S. aureus antigens, no. 18 (cross-react serologically with cell wall teichoic acid) and no. 46 of the reference pattern, were useful for serological diagnosis. The precipitin score, taking into account both the total number of precipitins and their titres, was the most useful diagnostic criterion. The present method can be used in selected cases to discriminate between S. aureus and non-S. aureus endocarditis. In cases with septicaemia only a positive test is of clinical value.

Adolescent↗
PubMed1978

The occurrence and nature of alveolitis-inducing substances in Aspergillus clavatus.

Five groups of antigens were identified in culture filtrates and extracts from spores and mycelia of Aspergillus clavatus fractionated by gel filtration, affinity chromatography, electrophoresis in polyacrylamide gels and chemical analysis. Some particulate and soluble fractions given by nasal inoculation provoked murine allergic alveolitis in non-sensitized and sensitized precipitin-negative, and sensitized, precipitin-positive, animals. Alveolitis-inducing substances appeared to be glycoprotein precipitinogens, which withstood proteolysis, were preferentially adsorbed by concanavalin A, but which were rendered almost inert by sodium periodate oxidation. Spore walls were particularly rich in allergenic substances extractable by alkaline hydrolysis. Delipidated dead spores provoked more severe disease in all immunological groups of mice than live spores. Polysacchraride extracts and acid hydrolysates of spore walls were unreactive.

Alveolitis, Extrinsic Allergic↗
PubMed1984

Detection of thyroglobulin in blood stains as an aid in the diagnosis of mechanical asphyxia.

Mechanical forces applied to the neck region are known to release certain amounts of thyroglobulin into circulation. In this experiment, an attempt was made to detect thyroglobulin in bloodstains as an aid in the diagnosis of mechanical asphyxia. Experimental bloodstains containing thyroglobulin at concentrations of 1, 2, 5 and 10 mu g/mL were prepared on a sheet of filter paper. Small pieces of bloodstains, measuring approximately 2.4 cm2 in area, were extracted with 0.1 mL of distilled water and the extracts were tested against an antihuman thyroglobulin serum by precipitation-electrophoresis. Bloodstains containing more than 1 mu g/mL of thyroglobulin formed distinct precipitin lines for up to one month of storage, while bloodstains containing more than 5 mu g/mL of thyroglobulin formed distinct precipitin lines for up to three months of storage. The present results suggest that the bloodstains can be utilized in the diagnosis of mechanical asphyxia.

Asphyxia↗
PubMed1982

Crossed-immunoelectrophoretic analyses of Trypanosoma cruzi epimastigote, metacyclic, and bloodstream forms.

Sonicated suspensions of epimastigote, metacyclic, or bloodstream forms of Trypanosoma cruzi were emulsified in Freund's complete adjuvant. Rabbits immunized with epimastigotes or metacyclics received five intramuscular (i.m.) injections of 1 x 10(9) sonicated trypanosomes at weekly intervals. Immunization with bloodstream forms included three i.m. injections of 5 x 10(7) and six injections of 2 x 10(8) sonicated trypanosomes. Selected antisera from these rabbits were employed in crossed immunoelectrophoretic studies against the homologous or heterologous extracts of sonicated trypanosomes. Extracts of epimastigote, metacyclic, and trypomastigotes produced 31, 29, and 11 precipitin peaks respectively against the homologous rabbit antisera. Tandem, crossed-immunoelectrophoresis of these extracts against antiepimastigote or antimetacyclic sera revealed that epimastigotes or metacyclics may each have at least four antigens that did not appear to be shared by the other, whereas each of these forms may have at least eight or nine antigens that were not detected with extracts from trypomastigotes. Cross-absorptions of antiepimastigote or antimetacyclic sera with live trypanosomes caused marked reductions in the numbers of precipitin peaks formed against the homologous extracts, but cross-absorptions with sonicated suspensions of epimastigotes or metacyclics showed that epimastigotes or metacyclics each have at least two antigens that were not detected in extracts of the other. Differentiation appeared to be accompanied by antigenic change. More antigens appear to be shared by epimastigotes and metacyclic forms than by trypomastigotes and epimastigotes or metacyclics.

Animals↗
PubMed1980

An assay for antibodies in leprosy sera reacting with ribonucleoprotein (RNP), a mycobacterial ribosomal antigen, using crossed immunoelectrophoresis with intermediate gel.

The technique of crossed immunoelectrophoresis with intermediate gel has been adapted to provide a quantitative assay for antibodies to a mycobacterial ribosomal antigen termed ribonucleoprotein (RNP) antigen. This antigen is no. 1 in the M. smegmatis reference system and corresponds to antigen no. 5 of M. leprae. The assay method measures changes in the rate of migration of the reference precipitin peak caused by the addition of serum in the intermediate gel and utilizes a lepromatous serum pool (LSP) both in the reference gel and as a standard in the intermediate gel. The anti-RNP activity in 24 leprosy sera differentiated the pauci-bacillary tuberculoid group (11 of 12 < 35% LSP) from the multi-bacillary lepromatous group (11 of 12 > 35% LSP). These findings confirm the work of others which indicates that assay of anti-RNP activity may have applications in the classification of leprosy patients and in the serodiagnosis of lepromatous leprosy infections. This method should also be applicable in other systems in which an antigen of high electrophoretic mobility forms a major precipitin in crossed immunoelectrophoresis.

Antibodies, Bacterial↗
PubMed1980

Efficacy of counterimmunoelectrophoresis in the detection of fungal antibodies in allergic bronchopulmonary mycoses.

The allergic bronchopulmonary mycoses (ABPM) are characterized by dual antibody response which includes immediate Type I and delayed Type III Arthus reactions to fungal antigens. In such cases the demonstration of precipitins is of diagnostic importance. In the present study the efficacy of counterimmunoelectrophoresis (CIE) has been compared with that of conventional agar gel double immunodiffusion technique (DID). Cases of allergic bronchopulmonary aspergillosis (ABPA) and candidiasis (ABPC) were diagnosed in a survey among patients with pulmonary diseases on the basis of positive clinical, radiological, mycological and immunological findings. Precipitating fungal antibodies were detected by CIE and DID techniques in 77 and 59 cases of ABPA and ABPC respectively. The former technique revealed double or multiple bands as against a single band observed in double immunodiffusion in a great majority of these cases. With CIE it was further possible to demonstrate the presence of precipitins in about 65 per cent of suspected cases of ABPM found otherwise negative in DID tests. The CIE has proved to be a more efficacious serodiagnostic procedure in allergic bronchopulmonary mycoses in which antibody levels are known to vary a great deal.

Antibodies, Fungal↗
PubMed1981

Heterogeneity of molecular size of factor VIII/von Willebrand factor in von Willbrand's disease.

Patterns of VIIIR:AG in the plasma and its fractions, cryoprecipitate and cryosupernatant, from various types of von Willebrand's disease (vWd) were observed by SDS 1.5% polyacrylamide gel electrophoresis - cross immunoelectrophoresis (SDS PAGE - CIE). VIIIR:AG in normal cryoprecipitate showed several precipitin peaks which correspond to molecular weights ranging from 8 X 10(5) to 1 X 10(7) daltons and are similar to those in normal plasma. Normal cryosupernatant VIIIR:AG gave smaller molecular weights from 8 X 10(5) to 2 X 10(6) daltons. VIIIR:AG in the plasma and cryoprecipitate from 2 patients with classical vWd gave low precipitin peaks with molecular weights in normal range. VIIIR:AG from 2 patients with subgroup A variant which showed fast anodal migration on the conventional CIE, presented 3 peaks with molecular weight of 8 X 10(5) to 3 X 10(6) which are similar to those in normal cryosupernatant. VIIIR:AG from 2 patients with subgroup B variant which showed normal migration on the CIE, gave normal patterns through all fractions.

Antigens↗
PubMed1981

Monospecific antibodies to rabbit lung ribonucleases.

Ribonucleases were isolated from normal rabbit lung by chromatographic techniques. Two active fractions were separated by elution from a sulfopropyl-Sephadex column with 0.2 and 1.0 M NaCl. These were further purified over 2000-fold from the original crude homogenate. Both RNase preparations showed more than one protein possessing RNase activity in polyacrylamide gel electrophoresis with either sodium dodecyl sulfate or low pH buffer systems. The purified fraction eluted at 0.2 M NaCl was injected into a goat. The resulting antiserum inhibited the RNase activity of both RNase preparations. Immunoelectrophoresis revealed three noncross-reacting precipitin arcs with the 0.2 M NaCl-eluted RNase and two noncross-reacting precipitin arcs with the 1.0 M NaCl-eluted RNase. Highly significant RNase activity was recovered from the beta arc in the former and the delta arc in the latter. By immunoabsorbant chromatography, the antiserum was separated into three monospecific fractions, anti-alpha, anti-beta, and anti-delta. Anti-beta and anti-delta inhibited, respectively, 60 and 33% of the 0.2 M NaCl-eluted RNase activity. Anti-delta inhibited 93% of the 1.0 M NaCl-eluted RNase activity. RNase activity in rabbit serum was not inhibited by any of the monospecific antibody fractions.

Animals↗
PubMed1982

Sperm specific antigen(s) in detergent extract of rhesus monkey spermatozoa.

Sperm surface antigens were prepared by detergent extraction (Hyamine-Triton, Rohm and Haas Co., Philadelphia, PA) of spermatozoa from rhesus monkeys. Heterologous antisera against the extracts were produced in female New Zealand White (NZW) rabbits by hyperimmunization. The antiserum was absorbed initially with sperm-free monkey seminal plasma and then with lyophilized tissue homogenates of liver, kidney, spleen, and pancreas from male rhesus monkeys. The unabsorbed antiserum produced at least three precipitin lines against detergent extract in double immunodiffusion tests and possessed sperm immobilization and sperm agglutination antibody activity. The absorbed antiserum showed one precipitin line against the detergent extract and retained sperm agglutination antibody activity only. The sperm agglutination antibody in the absorbed antiserum was completely removed by either epididymal or washed ejaculated rhesus monkey spermatozoa. The same results were obtained with ammonium-sulfate-precipitated immunoglobulin preparations. Immunoelectrophoresis revealed only one precipitation line with the absorbed immunoglobulin preparation. It is concluded that Hyamine-Triton extract of rhesus monkey spermatozoa contains a component which can be characterized as sperm-specific by immunoabsorption techniques and is apparently associated with sperm agglutination but not with immobilization.

Animals↗
PubMed1982

Schistosoma mansoni: fractionation of polysaccharide egg antigens by lectin affinity chromatography.

Crude polysaccharide antigen was extracted from Schistosoma mansoni egg homogenate by 44% aqueous phenol. The aqueous soluble polysaccharide extract was subjected to affinity chromatography with concanavalin A-Sepharose 4B. Two fractions (bound and unbound) were obtained; both of them gave precipitin lines with serum obtained from mice infected with S. mansoni. These precipitin lines gave partial identity. Further fractionation with wheat germ agglutinin-sepharose of the unbound fraction resulted in three antigenic fractions. These different antigens were eluted with different N-acetylglucosamine molarities (0, 0.05) and 0.5) and gave lines of identity when reacted against infected mouse serum. When the four antigenic materials were subjected to polyacrylamide gel electrophoresis and stained for proteins and polysaccharides no migration bands were observed. The chemical analysis of the two initial fractions showed a small percentage of amino acids in both fractions. Sugar analysis with gas-liquid chromatography showed different sugar composition of the two initial fractions.

Amino Acids↗
PubMed1982

Rapid serological diagnosis of Aspergillus fumigatus infection of the frontal sinuses and nasal chambers of the dog.

Aspergillus fumigatus precipitins in dog sera were detected by counterimmunoelectrophoresis (CIE). The procedure gave results within 90 minutes compared with 72 to 96 hours required in agar-gel double diffusion. Culture-filtrate antigens from two-week-old cultures of A fumigatus and two-day-old mycelial antigens produced the strongest reactions in CIE tests and the former antigen also revealed high titres in tests with serum from seven of 14 dogs with A fumigatus precipitins. Serum from these 14 dogs also reacted in CIE tests with a number of Penicillium species antigens.

Animals↗
PubMedDate not supplied

Serologic types and physiologic characteristics of 46 avian Pasteurella anatipestifer cultures.

Forty-six strains of Pasteurella anatipestifer isolated from different avian species were examined to determine their serologic types and physiologic characteristics. Serologic types were determined by a gel-diffusion precipitin test. Antigens from 39 field isolates reacted with antisera prepared from seven P. anatipestifer reference strains representing serotypes 1, 2, 4, 6, and 7. Antigens from five isolates did not react and could not be typed with available reagents. Gel precipitin reactions involving serotype 1 (43.6%) and serotype 2 (25.6%) were the most prevalent. Generally, the physiologic characteristics from 40 tests were typical for P. anatipestifer, and variations were observed among the strains in urease production, hemolysin production, litmus milk reaction, and gelatin liquefaction.

Animals↗
PubMed1976

The serological specificities of Salmonella typhi antigens.

Sera prepared with two different strains of Salmonella typhi were analysed against all the soluble antigens isolated from S. typhi 0901, S. typhi Ty2 and S. typhi Vi. Agar-gel diffusion against individual sera showed that, in all the sera, antibodies were induced against somatic antigens and free proteins. Absorptions of the sera with polysaccharides, split from the somatic antigens, removed the antibodies induced against the polysaccharide and its proteinic carrier in most of the somatic antigens of S. typhi 0901. The antibodies left in the absorbed sera reacted against the proteinic moieties of more complex somatic antigens of S. typhi and against free proteins from all the analysed strains. Only the absorption with proteins removed all the precipitating antibodies from the sera. Moreover, in incomplete absorptions with proteins, the first antibodies removed are the antipolysaccharides, since antibodies are never induced against the haptenic polysaccharide but against somatic conjugates; in these the proteinic moiety eventually varies with every batch of bacteria. The sera exhausted of precipitins still agglutinate the bacteria, thus confirming the assumption that agglutinins and precipitins may be different antibodies.

Animals↗
PubMed1981

[Humidifier lung].

Breathing air from a humidifier or an air conditioning unit contaminated by various microorganisms can cause an acute lung disease involving fever, cough and dyspnea, termed "humidifier fever". This type of hypersensitivity pneumonitis was first described in 1959 by PESTALOZZI in the Swiss literature and subsequently by BANASZAK et al. in the Anglo-American. Here a chronic form of this disease which led to pulmonary fibrosis is described: A 37-year-old woman who works in a cheese shop presented with dyspnea which had been progressive over two years, weight loss, a diffuse reticular pattern radiographically and a severe restrictive defect in lung function tests. Open lung biopsy revealed chronic interstitial and alveolar inflammation with non-caseating granulomas and fibrotic changes. Circulating immune complexes and precipitins against the contaminated humidifier water and cheese mites were found, but no antibodies suggesting legionnaires' disease. Two out of five otherwise healthy employees of this cheese shop, where a new humidifying system had been installed 7 years earlier, also had precipitins against the contaminated water from the humidifier and the cheese mites. Despite ending of exposure and longterm steroid and immunosuppressive therapy, the signs and symptoms of pulmonary fibrosis persisted. Contrary to the acute disease, this chronic form is termed "humidifier lung". The importance is stressed of investigating the possibility of exposure to contaminated humidifiers or air conditioning units in all cases of newly detected pulmonary fibrosis.

Adult↗
PubMed1980

Potential health hazards of the water resources development: a health survey in the Phitsanulok Irrigation Project, Nan River Basin, Northern Thailand.

A health survey was carried out among residents of 33 villages under the Phitsanulok Irrigation Project Area, Nan River Basin, Northern Thailand, whereby general health conditions were examined including intradermal tests for schistosomiasis japonica, stools for intestinal parasites and sera tested by circumoval precipitin test for antibodies to S. japonicum and by agglutination test for leptospiral infection. Health investigations revealed that 913 (60.9%) of 1,499 people examined had experienced gastro-intestinal disorders, 254 (17%) had poor oral hygiene and a few had mild anemia. 799 (62%) of 1,298 examined had intestinal parasites. Hookworm (45%) was most common, followed by Opisthorchis viverrini (20%), Entamoeba coli (10%), Ascaris lumbricoides (6%), intestinal flukes (2%), Enterobius vermicularis (1%) and Giardia lamblia (1%). 149 (10%) of 1,422 people gave positive skin reaction to crude S. japonicum antigen. Circumoval precipitin test was strongly positive in 7 (1.6%) persons out of 449 tested for schistosome infection and 6 (4.0%) out of 1,358 people were positive for leptospiral infection. The significant endemic diseases as potential health problems in this water resources development are discussed.

Adolescent↗
PubMed1980

Analyses of some mite antigens found in house dust and in food material.

Antigen analyses by micro-Ouchterlony, immunoelectrophoresis and passive hemagglutination tests were carried out on extracts from each of the following cultured mites: Dermatophogoides farinae, Dermatophogoides pteronyssinus, Tyrophagus putrescentiae, Aleuroglyphus ovatus and Carpoglyphus lactis; and on an extract from the culture medium for each mite. Antisera were raised against each extract or each medium in rabbits or in guinea pigs. Up to 5 precipitin lines by micro-Ouchterlony test and up to 8 precipitin arcs by immunoelectrophoresis were detected between an extract and its homologous (corresponding) antiserum. Some of the lines or arcs were seen consistently to be common to all species tested. The absorption of common antigens in an extract by heterologous (noncorresponding) antiserum was employed to find the antigens specific to D. farinae and D. pteronyssinus. By these tests, at least 3 species-specific antigens for D. farinae and 2 for D. pteronyssinus were found. In passive hemagglutination tests, higher titers were obtained mainly in homologous systems, although weak cross reaction was observed in every system. Culture media did not participate in any reaction combinations used here and did not interfere with the analyses.

Allergens↗
PubMed1994

Antibody response in rabbits infected with Rochalimaea henselae, Rochalimaea quintana and Afipia felis.

Antibody responses in three pairs of rabbits inoculated with live Rochalimaea henselae, Rochalimaea quintana and Afipia felis were studied by enzyme immunoassay with whole-cell and bacterial sonicates as antigen. No differences in measured antibody responses were found with the two types of antigen preparation. Two rabbits did not respond with antibody production. In the remaining rabbits there was a low-titred antibody response that showed no significant cross-reaction with related bacteria. After rechallenge the antibody response rose significantly and there was significant cross-reaction with related bacteria. The antigens involved in the antibody response were examined by crossed immunoelectrophoresis. After the initial inoculation 5-7 precipitin lines of the reference diagrams were deflected, including lines which cross-reacted with antigens found in related bacterial species. After reinoculation several more precipitin lines were deflected, including additional lines cross-reacting with antigens present in related bacteria and common bacterial antigens.

Animals↗