POSSIBLE ROLE OF PRECIPITINS AGAINST ASPERGILLUS FUMIGATUS.
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The colloidal dye immunofiltration assay (CDIFA) combines the concepts of the double-antigen sandwich assay, the Dot immunofiltration assay, and colloidal dye-linked antigen technique to produce a new dye immunoassay for antibody detection in schistosomiasis. The CDIFA consisted of soluble egg antigen (SEA) of Schistosoma japonicum coated onto nitrocellulose membrane, mounted on a flow-through test device to provide the assay capture matrix. SEA absorbed to a red colloidal dye, R-3, produced in China, served as the antigen-antibody complex detecting reagents. The results showed that the sensitivity of the CDIFA was 100% in 35 cases of acute schistosomiasis (35/35), 98% in 50 cases of chronic schistosomiasis (49/50). The specificity of the assay was 99.4% in 180 healthy individuals (179/180). The cross-reaction was 13.3% in 30 cases of paragonimiasis, 2.6% in 38 cases of clonorchiasis sinensis and 0% in 20 cases of hookworm infection, 20 cases of fasciolopsiasis and 16 cases of ascariasis. The results were similar to those detected by routine enzyme-linked immunosorbent assay (ELISA). In a field evaluation of the CDIFA kit, the positivity rate of the CDIFA was 97.5% in 157 cases of schistosomiasis, compared with 91.1% with the circumoval precipitin test (COPT). The dye-labeled SEA conjugate was stable at room temperature for at least 6 months. The results indicated that the CDIFA provided an economic, simple, rapid, robust test for the detection of schistosome infection, suitable for a wide variety of field applications without any instrumentation.
An indirect solid phase radioimmunassay (SPRIA) performed in flexible polyvinyl microtiter plates was modified for quantitative determination of antibodies to various bacterial antigens. Parameters affecting quantitation of the assay were investigated by using meningococcal serotype antigens and rabbit antisera plate was essentially complete after 1 hr at 37 degrees C, and only 4 to 8% of the bound antigen was released during the course of the assay. About 90% of the specific primary antibody (PAb) added to the antigen-coated wells was bound after overnight incubation at room temperature, whereas, 10 to 25% of the bound PAb was released before completion of the assay. Under conditions of limiting PAb the time required for saturation binding of 125I-anti-immunoglobulin (secondary antibody = SAb) to the PAb was dependent on the concentration of SAb. At a concentration of 20 ng SAb/25 mul, maximum binding occurred in 12 to 16 hr at 22 degrees C. Uncer conditions of extreme SAb excess the cpm of 125I-SAb bound was directly proportional to the amount of PAb bound. Under these conditions the cpm of 125I-SAb bound per well can be related to the amount of PAb added per well by a time-dependent calibration coefficient K(t) which is the product of three parameters: 1) the specific activity of the 125I-SAb, 2) the ratio of PAb bound to SAb bound and, 3) the fraction of added PAb remaining bound to the plate. Experimentally determined values for these parameters were used to calculate K(t) and quantitate the specific antibody in nine rabbit antisera with the SPRIA. A close correlation (r=0.985) was found between these results and the results of quantitative precipitin tests performed by using the same antisera and antigens. Although the SPRIA results were an average of 33% lower, a more accurate value for K(t) can easily be determined by performing the SPRIA on several sera calibrated by the quantitative test. Reproducibility of the SPRIA for 10 replicate determinations was +/- 6.7%. The assay described is capable of measuring a minimum of about 0.5 mug antibody/ml of serum and appears to be applicable to many different antigens.
By a case of bird fancier's lung proved serologically by precipitin tests the importance of a detailed case history is being stressed since there is an increased frequency of exogenous allergic alveolitis. The paper shall contribute to an early diagnosis to avoid irreversible professional and non-professional damages. For all cases of manifest disease constistent avoidance of allergens is demanded.
The response of precipitin-negative sera from non-selected asthmatic patients against a range of somatic and culture filtrate antigens of Aspergillus fumigatus in ELISA for anti-A. fumigatus IgG is described. Antibody to the various antigens was widely distributed but the precise distribution was dependent upon antigen type. To determine how the selection of negative reference sera from precipitin-negative sera can influence the discrimination between precipitin-positive and -negative sera in ELISA, panels of 10 sera were chosen from the extremes of the frequency distribution and by random selection. The closest agreement between precipitin testing and ELISA was seen when randomly-selected negative reference sera were incorporated in the assay, although the degree of correlation was dependent upon the detector antigen. These findings demonstrate the requirement for careful selection of sera included as negative reference sera in ELISA for anti-A. fumigatus IgG.
The current study summarizes our findings of anti-schistosome egg antibody by the circumoval precipitin test for two different populations in Puerto Rico. One group, exclusively males more than 40 years of age and from all municipalities on the island, was from the Veterans Administration Hospital for the period 1988-1997. The second group resided southeast of San Juan, around the municipality of Caguas and adjacent municipalities east of Caguas, was of both sexes and mostly until 1997 of undetermined ages for the period 1993-1997. Results reveal a yearly decrease in testing requests from the Veterans Administration Hospital from 148 in 1988 and 1989 with 16% positive to three in 1996 through 1998 with none positive. This decrease in testing requests was because of a decrease of suspicion of schistosomiasis in this group. The other patient population from the Caguas region showed a gradual but continuous decrease in seropositive individuals from 21% in 1993 to 12% in 1996, with precipitous decrease to 5% in 1997 and only 1% in 1998. Moreover, there were four patients from which at least two serum samples were obtained one or two years apart and tested. In each instance the more recently obtained sample had lower antibody reactions than the first as reflected in lower percentages of positive egg reactors. The fact that they were treated with praziquantel after the first testing also suggests that the infected population was being eliminated through chemotherapy. These combined results suggest the elimination of infections with Schistosoma mansoni in the traditionally high prevalence regions east of San Juan in the absence of any proactive control efforts in Puerto Rico. Because of the rapid urbanizing of Puerto Rico, the one identifiable control effort is economic development and well being.
The group antigens of streptococcal groups P and U were extracted with formamide and purified on diethylaminoethyl-Sephadex A-25 and Sephadex G-200. The antigens were shown to be polysaccharides located in the cell walls of the organisms. In a precipitin test, the P and U group antigens did not cross-react with homologous sera of each other, nor with specific antiserum of the group antigen of group E streptococci. The polysaccharide comprising the group P antigen contained rhamnose, glucose, galactose, glucosamine, and galactosamine; the group U antigen was similar in composition but lacked galactosamine and contained more galactose.
Various parameters of replication of Marek's disease virus (MDV) were studied in cells derived from genetically resistant (line 6) and susceptible (line 7) chickens. Cell-free and cell-associated preparations of three strains of pathogenic MDV (JM, GA, and Id-1) replicated equally well in cells of resistant and susceptible chickens. There were no demonstrable differences between the two genetic sources of cells for the chronological appearance and type of cytopathic effects and for the type of virus-induced antigens detectable by immunofluorescent and agar-gel precipitin tests. MDV propagated for about 4 weeks in cells of resistant line 6 chickens remained nonvirulent for line 6 and fully virulent for line 7 chickens. The lack of expression of genetic resistance to Marek's disease at cellular level contrasts the infection mechanisms of MDV from those of viruses associated with lymphoid leukosis in chickens.
The aim of this work was to verify the colonization of Panstrongylus megistus on artificial ecotopes in Florianópolis, in the Santa Catarina Island, South Brazil. For this purpose 443 houses of the Lagoa district and 779 house annexes (524 chicken-houses, 46 corrals and 209 storage-houses) in 9 different places were examined from 1985 to 1992. These ecotopes, which include ceilings and basements, were checked after application of dislodging liquid (Pirisa 5%). Colonization by P. megistus was verified in two houses, three chicken-houses and one storage-house of the Lagoa district, where eggs, nymphs and adults were collected. To verify local reports of P. megistus occurrence, another two houses and one school were investigated. The colonization at all of these places was confirmed. In the 9 artificial ecotopes examined, 559 eggs, 305 nymphs and 24 adults were collected. The infection rate of P. megistus by Trypanosoma cruzi was 55.3% (182/329). A similar infection rate of 56.5% (78/138) was obtained in adults of P. megistus from sylvatic ecotopes and in adults captured in the houses by the inhabitants between 1983 to 1991. Precipitin tests revealed blood from just one source in 94.0% of the insects (170/181). Human blood was found in 80.6% (25/31) of the adults and in 5.8% (1/17) of the nymphs captured in the houses. These results suggest the need to ally serious epidemiologic vigilance to the effort of the inhabitants in order to avoid the risk of domiciliation of P. megistus in the houses.
A solid-phase radioimmunoassay (RIA) was developed for schistosomiasis japonica using extracted egg antigens and compared with circumoval precipitin test (COPT) results on 20 sera from known S. japonicum-infected individuals and on 10 control sera. The quantitative RIA very clearly differentiated between infected and uninfected individuals with highest titers being obtained in teenagers. However, in the series employed, information relevant to immunodiagnosis of S. japonicum infection was contained in the non-quantitative but simple COPT and little was apparently added to the quantitative but expensive RIA.
This paper deals with the fluorescent antibody (FA) method for identifying six commonly occuring and two rare groups of streptococci by using commercially prepared (Difco) conjugates. We have shown that group-specific FA produced frequent cross-reactions with heterologous groups of organisms. These reactions varied with different strains of the same serogroup. Nonetheless, there was distinct overall patterns in the intensity and appearance of the homologous and heterologous reactions. When monitored by the precipitin test with Rantz and Randall extracts, these patterns led to the correct identification of 90 to 100% of specimens of serogroup A, B, C, and G streptococci. Many members of groups D and F also showed distinctive reaction patterns. However, there was a significant number of strains of both groups D and F that either failed to strain or stained poorly with the homologous conjugate. As a result, the identification of these serogroups by FA was less reliable.
Intact bacterial membranes have been isolated from protoplasts prepared from Group A streptococci by a cell wall-dissolving enzyme. A membrane fraction with identical chemical and serological properties has been obtained by differential centrifugation of mechanically disrupted streptococci. The membrane is chemically distinct from the cell wall and is composed of 72 per cent protein, 26 per cent lipid, and 2 per cent carbohydrate. Capillary precipitin tests and analysis by microdiffusion have demonstrated that the membrane contains antigens distinct from those of the cell wall and from those of the cytoplasm which it envelops. Evidence is presented which demonstrates that this antigenic material is common to the membranes of Group A streptococci, and that it can be distinguished by immunodiffusion from related antigenic substances present in membranes of several other serological groups of hemolytic streptococci. This antigenic material does not cross-react with the membrane antigens of other Gram-positive cocci.
The capillary tube precipitin test was used to determine the host utilization patterns of Culiseta inornata in southwestern Manitoba. Ruminant blood was identified in 83.3% and equine blood in 15.8% of 1,036 positively reacting blood-meals. Human, swine and avian blood accounted for 0.9% of these blood-meals and mixed blood-meals accounted for 1.5% of the total. Culiseta inornata preferentially fed on large mammals, and selection between cattle and horses reflected the relative abundance of these two hosts rather than a specific preference for either one.
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Sera frour 146 patients with malignancy, 59 normal controls and 42 patients hospitalized with non-malignant diseases were examined by a precipitin test with monoclonal rheumatoid factor (mRF) for the presence of circulating immune complexes containing IgG. Forty-two (29%) of the sera from cancer patients but only two of the sera from patients in each of the control groups contained such material. Similar results were obtained with a radioimmunoassay for immune complexes based on the same mRF. Sera from 23 of 65 patients with metastatic malignancy (35%) had elevated levels of immune complexes by this latter test. The presence of the material was not related to the source of malignancy, presence of carcinoembryonic or hepatitis antigens or of such autoantibodies as rheumatoid factor or anti-DNA. By density gradient ultracentrifugation the reacting material was identified as being of molecular size 19s or greater. It has not yet been further characterized with regard to the nature of any antigens present.
Field experiments were made in southern England to re-examine the possibility that mosquitoes in Britain might feed on wild rabbits and hence be vectors of myxomatosis. Mosquitoes of several species were attracted to rabbits enclosed in cylindrical traps and in a trap in which the animal was placed in a wire mesh cage. Substantial numbers of mosquitoes were also caught biting, or attempting to bite, tethered rabbits which were not in cages or traps. Evidence that mosquitoes fed on wild rabbits under natural conditions was obtained from results of precipitin tests made on blood-smears collected from mosquitoes caught resting amongst vegetation. On a few evenings mosquitoes were seen to be attracted to healthy wild rabbits and apparently attempting to feed on them. Batches of two mosquito species collected from the field were infected with myxoma virus.It was concluded that contrary to previous beliefs mosquitoes in Britain feed to a certain extent on wild rabbits, and therefore are potential vectors of myxomatosis. No attempts were made to assess their relative importance in the transmission of the disease, which in Britain is transmitted mainly by the rabbit flea.
Ten monoclonal antibodies (McAbs) raised to Schistosoma japonicum eggs could be assigned using several serological and immunochemical techniques to 3 groups. The McAbs, termed A, B and C-McAbs, apparently recognize carbohydrate epitopes that can be located on the same antigen molecule. The antibodies, generally of IgM isotype, are idiotypically related. They are distinct from another IgM McAb (Group D-McAb) the carbohydrate target epitope of which can also be associated with the epitopes of A, B and C-McAbs. The McAbs produce large vacuolated bleb reactions in the circumoval precipitin test (COPT) and target epitopes have different representations in various life cycle stages such as immature and mature eggs, male and female worms (including S. mansoni). Antigens affinity purified on columns containing A, B, C and D-McAbs stimulate proliferation of T cells from egg-sensitized mice and elicit DTH reactions in such mice. This raises the possibility that the target antigens of these carbohydrate-reactive monoclonal antibodies are immunopathologic and involved in egg-induced granuloma formation.
A survey of the species composition and distribution of the Culicoides midge populations at a range of sites where bluetongue is enzootic isolated a group of dominant species: C. cornutus, C. grahamii, C. magnus, C. milnei, C. pallidipennis and C. 23. dagger Monthly light-trap sampling of Culicoides showed that the population densities of the dominant species greatly increased after the rain seasons and that these species concentrated around flocks of sheep and cattle. The larval habitats of C. cornutus and C. pallidipennis were found associated with stock pens. Precipitin tests on blood-fed Culicoides showed that most of the dominant species regularly feed on sheep and cattle. Bluetongue virus was isolated from C. milnei, C. pallidipennis and C. 23. Serological surveys of wild and domestic bovids from the enzootic area showed a high proportion with antibody to bluetongue virus. The colonization of C. cornutus, a potential vector, is described briefly. A causal relationship between peak rainfall in April-May, peak numbers of Culicoides in May-June and peak bluetongue incidence in June-July is postulated. The vector status of the above species and C. austeni was evaluated.