The use of the latex agglutination reaction for the diagnosis of a Brucella infection

The authors have developed the optimum conditions for the preparation of antigenic diagnosticum based on latex manufactured in the USSR. To sensitize latex with the diameter of microspheres equal to 0.83 microns, Brucella polysaccharide was used in a dose of 100 micrograms/ml. As stabilizer, polyvinylpyrrolidone at a concentration of 0.1% was used. The specificity and sensitivity of the diagnosticum were studied in analysis of serum samples taken from 102 healthy donors and patients with infectious diseases of nonbrucellar etiology and from 120 patients with different forms of brucellosis. The specificity of the diagnosticum was found to be 94.1% and its sensitivity, 77.5%. Comparative study of the latex agglutination test with other serological tests showed that the former test is highly effective both in acute and chronic forms of the disease. A high degree of correlation between the agglutination test, Coombs' test, the passive hemagglutination test and the latex agglutination test was established (r = 0.83, 0.72 and 0.62, respectively).     

Development of antilipase immunoglobulin polymer diagnosticum for the detection of Vibrio cholerae eltor, possessing hemolytic and lipase activity

The possibility of using a heterogeneous, but structurally similar antigen--the commercial preparation of Pseudomonas sp. lipase (Sigma, USA)--for the development of polymer diagnosticum aimed at determination of lipase production in cholera vibrios was shown. The new diagnosticum (antilipase antibodies) on a polymer carrier was used in the serological volume agglomeration test for the detection of hemolytic atoxigenic V. eltor, obtained from environmental, objects, which produced lipase in 80% of cases. The differentiating capacity of the diagnosticum was confirmed on 120 V. eltor cultures isolated from environmental objects. The newly developed diagnosticum makes it possible to determine the lipase activity in cholera vibrios of different biovars and serovars.     

The results of a controlled field trial of a dried erythrocytic immunoglobulin diagnostic agent for detecting the hepatitis A virus antigen]

The results of the controlled field trial of lyophilized erythrocytic immunoglobulin diagnosticum for the detection of hepatitis A virus antigen in the urine and feces of patients are presented. This diagnosticum was used for the study of urine and fecal samples from 225 patients (of these, 176 had hepatitis A) and 54 healthy persons in the passive hemagglutination (PHA) test. Their blood sera were studied in the PHA test (to detect HBsAg) and the radioimmunoassay (to detect anti-HAV IgM). The immunoglobulin diagnosticum under study was found to be nonspecific and faintly sensitive and, therefore, unsuitable for use in medical practice.     

The preparation of an IgG diagnostic agent based on stained polyacrolein latexes for use in the latex agglutination reaction

IgG diagnosticum for measuring the concentration of 131I-labeled IgG antibodies to enteric antigen beta 1MA by the latex agglutination inhibition (LAI) test has been prepared on the basis of polyacrolein latexes. A method for the titration of anti-IgG antibodies with the use of the above diagnosticum has been developed, based on the late, agglutination (LA) test. The optimum conditions for the microtitration variant of the LA and LAI tests have been defined. High sensitivity, specificity and simplicity of analysis with the use of latex IgG diagnosticum have been demonstrated. The newly developed methods have been successfully used in laboratory trials of a new diagnostic radiopharmaceuticals for the assay of 131I-labeled antibodies in this preparation and for the detection of side effects of immunization on the recipients.     

Results of a trial of a plague monoclonal antibody erythrocyte diagnostic agent

Plague antibody monoclonal erythrocyte diagnosticum was studied in serological tests simultaneously with commercial plague antibody erythrocyte diagnosticum prepared on the basis of hyperimmune horse serum and with commercial plague antigenic erythrocyte diagnosticum. In this investigation the suspensions of numerous strains of Yersinia pestis, other closely related and heterologous organisms, experimentally infected wild and laboratory animals, as well as samples of materials obtained from small rodents caught in several natural foci of plague, were studied. The monoclonal diagnosticum was, practically, not inferior to the similar commercial preparation with respect to the frequency of positive results and the activity of the materials under study in serological tests, but showed greater specificity, as it reacted strictly with Y. pestis capsular antigen.     

Diagnostic suspension for detection of the infective agent of tularemia

The matrix for obtaining silica-based diagnosticum was selected, its activation with surfactant was optimized and its immobilization with tularemia immunoglobulins was carried out. In the glass suspension agglutination test (SAT) the sensitivity of the diagnosticum was 3.125 x 10(6) to 6.25 x 10(6) microbial cells per ml; the duration of SAT, including the evaluation of its results, was 1-5 minutes.     

Use of the pneumococcal serovar 3 erythrocyte antibody diagnostic agent for analyzing polysaccharide-containing preparations

Antibody diagnosticum to pneumococci (serovar 3) has been prepared. The analysis of different antigenic preparations of pneumococci (serovars 1,3 and 6B) and the filtrates of culture fluid obtained in the process of the cultivation of bacteria belonging to serovars 3,9N and 23F has been carried out by means of the indirect hemagglutination test. The new diagnosticum has proved to be type-specific. This diagnosticum can be used for the evaluation of the quantitative content of specific polysaccharide in different antigenic preparations of serovar 3 pneumococci, as well as for the determination of the content of specific polysaccharide directly in the culture fluid, which constitutes a step in the determination of the period of the maximum synthesis of polysaccharide, necessary for the optimization of the process of the cultivation of pneumococci.     

The choice of rational methods for determining antibodies to Yersinia enterocolitica. The agglutination reaction

Six tests suitable for the detection of antibodies to Y. enterocolitica, serovars 03 and 09, were studied. The results of the study of hyperimmune sera revealed that the agglutination test and the indirect hemagglutination test were the most promising methods and deserved efforts for their further development. Bacterial diagnostic agents were prepared from live, heat-killed and formalin-killed Yersinia cultures and tested. The titration of homologous and heterologous sera revealed no essential differences in the preparations; in the serological examination of healthy persons (102), patients with Yersinia enteral infection (150) and with other acute enteral infections (92), the results obtained with the use of the above-mentioned preparations carried the highest information content. OH diagnostic agents proved to have the highest specificity; of these, the diagnosticum obtained by treatment 0.3-0.4% formalin vas found to be the most stable and technologically effective preparation. The minimal diagnostic titer of the agglutination test for the presence of Yersinia enteral infection was established (1:60 = 1:320). This titer was determined in 73% of the patients with bacteriologically confirmed Yersinia enteral infection and only in 1% of healthy persons and patients with other acute enteral infections.     

Myxovirus Dissemination by Air

Myxoviruses including 150 strains of parainfluenza 1, 15 of parainfluenza 3 and five of influenza B virus were isolated from nasopharyngeal secretions obtained from 300 children less than 3 years of age who developed acute laryngotracheobronchitis during the preceding 48 hours. The patients were examined between October 1966 and January 1967, the peak monthly rate of virus isolation (67%) occurring during January. Parainfluenza 1 virus was isolated from air obtained in the vicinity of one of 30 children whose nasopharyngeal secretions yielded this agent. Samples comprising 150 litres of air were collected for virus assay by placing an Andersen sampler about 60 cm. from the child''s face inside an oxygen tent which surrounded the patient. These findings confirm previous observations that parainfluenza 1 virus is the dominant agent associated with acute laryngotracheobronchitis in children in Toronto, and they show that this virus is disseminated in the air.     

The diagnostic value of a meningococcal-B erythrocyte diagnostic agent based on the group-specific polysaccharide

In this work the diagnostic value of group B meningococcal erythrocyte diagnosticum was determined. 585 blood serum samples taken from adult donors were studied: 220 samples from practically healthy persons and 365 samples from 144 patients with meningococcal infection and purulent bacterial meningitis of nonmeningococcal etiology. Group B meningococcal erythrocyte diagnosticum was found to possess serological activity and to reveal the growth of specific antibodies in the sera of patients with meningococcal infection, serologically confirmed by the isolation of group B meningococcal culture, in 100% of cases on weeks 2-3 of the disease. Diagnostic characteristics--specificity and sensitivity--for group B erythrocyte diagnosticum were, respectively, 90.2% and 63.5%. The study revealed that antibodies to several group-specific meningococcal polysaccharides in blood sera can be simultaneously determined in the passive hemagglutination test with a set of erythrocyte diagnostica, which should be taken into consideration in the clinical interpretation of serological results.     

Observations on Clinical and Immunofluorescent Diagnosis of Parainfluenza Virus Infections

Immunofluorescent techniques have been applied to nasopharyngeal secretions for the rapid diagnosis of parainfluenza virus types 1, 2, and 3 infections. Seventy-five infections were found by isolation techniques; 55 of these had nasopharyngeal secretions taken and 53 were positive by direct examination. A comparison of the results of 60 neutralization tests with immunofluorescence applied to monkey kidney isolations showed complete agreement. Immunofluorescence appeared to be a satisfactory method for differentiating the various haemadsorption viruses. The importance of parainfluenza viruses and respiratory syncytial virus in croup was noted and the association of the parainfluenza viruses with acute respiratory virus infection was confirmed. The clinical relationship between respiratory syncytial virus and parainfluenza virus type 3 is discussed.     

The neutrophil receptor apparatus in typhoid fever

In 157 typhoid fever patients and 36 practically healthy persons the content of neutrophils forming complement-dependent rosettes (NEAC rosette-forming cells), as well as rosettes with sheep red blood cells (NE rosette-forming cells) and with typhoid erythrocyte diagnosticum (NS rosette-forming cells), has been studied. The data obtained in this investigation indicate that antigen-binding neutrophils play an active functional role in the pathogenesis of typhoid fever at its acute stage. The values characterizing the content of N rosette-forming cells may be used as a criterion for the evaluation of the severity of infection, as well as for the prognostication of complications and relapses.     

Erythrocyte diagnostic kits for detection of Pseudomonas aeruginosa exotoxin A

The use of formulated chick red blood cells loaded with IgG preparations and affinity-purified antibodies, in comparison with initial immune serum to P. aeruginosa exotoxin A (ETA), has been shown to increase the sensitivity of antibody erythrocyte diagnosticum (AbED) 17-fold and to ensure the detection of ETA at a concentration of 1.2 mg of protein per ml. The passive hemagglutination (PHA) test with AbED has proved to be a more sensitive method for the detection of ETA than the antibody neutralization test with the use of antigenic erythrocyte diagnosticum, the latex agglutination test, the coagglutination test and the enzyme immunoassay. The PHA test has permitted the detection of ETA in the culture fluid of 80% of P. aeruginosa cultures under study.     

A method for the passive agglutination of polymer dispersions for the diagnosis of legionellosis]

The method of synthetizing dispersions of gelatin-modified polyacrolein microspheres 1.5-2.5 micrograms in diameter, used as a solid-phase carrier for the preparation of immunodispersion diagnostica, has been developed. The possibility of using immunodispersion diagnosticum in the passive agglutination test with hyperimmune rabbit sera has been demonstrated. High activity and specificity of immunodispersion diagnosticum, combining methodological simplicity and rapidity characteristic of agglutination, has been shown.     

The rapid diagnosis of acute streptococcal infection in children]

The present work deals with trials of the method of rapid diagnosis of streptococcal infection, carried out in children's infectious hospital, with the use of a new diagnostic kit. The proposed diagnosticum has proved to be highly sensitive and specific in scarlet fever and tonsillitis. The sensitivity and specificity of the diagnosticum depend on the duration of the disease, prehospital treatment and the quality of the bacteriological analysis.     

Obtaining erythrocyte diagnosticum for the detection of chlamydial antigen

It is shown possible to obtain erythrocyte diagnosticum for detection of chlamydial antigen, whose cell basis consists of a formalinized suspension of ram erythrocytes, sensibilized with hyperimmune antichlamydial sera by means of the amydol. High sensitivity and specificity of the diagnosticum, absolute correlation with the data obtained in the complex examination of patients with the urogenital tract pathology of the chlamydial etiology by other methods were determined in the course of investigation in the indirect hemagglutination test with diagnosticums of scrape specimens from these patients.     

Occurrence of Helicobacter pylori specific antigens in diseases of gastrointestinal tract

The results of the immunophoretic analysis of H. pylori antigenic structure are presented. Along with H. pylori O-antigen, specific surface antigen of acidic nature, relatively thermolabile with characteristics similar to those of K-antigens, was detected. A diagnosticum based on the coagglutination test for the rapid detection of H. pylori specific antigens in the patients' biological fluids (coprofiltrate, saliva) by the noninvasive method was developed. The circulation mode of H. pylori in human body and its correlation with the clinical symptoms of the disease was shown. H. pylori antigens were very frequently detected (in 65 - 83% of cases) in patients with acute bacterial enteric infections: shigellosis, salmonellosis, yersiniosis, campylobacteriosis. The newly developed method seems to be promising for the rapid diagnosis of H. pylori associated infections, for the prolonged monitoring and evaluation of the effectiveness of antibacterial therapy.     

Application of diagnosticum based on functionalized carbon nanoparticles for monitoring of immunoglobulins affinity purification

A diagnostic reagent based on carbon nanoparticles covalently functionalized with streptococcus G protein was applied to construct a system for monitoring and optimization of the technology of affinity purification of rabbit polyclonal antibodies to alpha-fetoprotein. The developed system allows a short-time (45 min) semiquantitative assessment of the immunoglobulins (IgG) of most higher animals and human beings in blood serum samples and eluate. IgG detection sensitivity using the carbon-G-protein diagnosticum was 80 ng/mL. Approaches to stabilizing the components of the analysis system, which ensures the preservation of their functional properties during long storage, were developed. The storage life of the diagnosticum was more than 20 years, and that of immunosorbents was more than year and a half. A technique of long-term immunosorbent storage was developed. Application of the developed test-system do not require registration equipment.     

The role of respiratory tract viruses in the etiology of obstructive bronchitis in infants

Out of 524 children with acute respiratory infections in 141 obstructive bronchitis was diagnosed (OZO). Seventy cases could be linked to viral infection. Viral infections tested (influenza virus A, B, parainfluenza typ 1-3, RSV, adenoviruses) were more frequently associated with OZO than other acute respiratory infections of unknown etiology. Majority infections induced by influenza virus A and parainfluenza virus typ 2 were accompanied by OZO symptoms. Of the highest risk of acquiring OZO despite of viral infection participation, were children of 4-12 months of age. OZO associated viral infections prevailed during autumn-winter season, while in spring-summer period undetermined factors were the major cause of OZO. In serum samples of children with OZO, despite of etiology of the disease, higher level of IgE was found than in a group of children without the symptoms. In the case of OZO of unestablished etiology the level of serum IgE was significantly higher than in the cases when viral etiology of the disease was found.