[Seroimmunologic monitoring of microorganisms of Rickettsia and Bartonella species microorganisms in the Moscow region]. / Seroimmunologicheski monitoring mikroorganizmov rodov Rickettsia i Bartonella v Moskovskom regione.

Serological study of 788 blood sera, taken from residents of the Moscow region was conducted using antigens of microorganisms of the genera Rickettsia and Bartonella. The first group under examination consisted of 355 patients with diagnosed diseases of nonreckettsial nature. The second group includes 433 healthy adults working at a meat processing and packing factory. The main method used for sera survey was the indirect immunofluorescence test. In the sera taken from the first group of subjects specific antibodies to R. prowazekii, R. typhi, B. quintana, B. henselae antigens were detected in 2.3%, 5.1%, 4.0% and 2.9% of serum samples respectively. In the serum samples taken from the second group the proportion of antibodies to R. prowazekii, R. typhi, B. quintana, B. henselae antigens was different: 0.5%, 3.3%, 1.7% and 4.0% respectively. In total, specific antibodies to R. typhi and B. henselae prevailed over specific antibodies to R. prowazekii and B. quintana twofold.

[The use of the latex agglutination reaction for the diagnosis of a Brucella infection]. / Ispol'zovanie reaktsii aggliutinatsii lateksa dlia diagnostiki brutselleznoi infektsii.

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).

[Use of the ELISA immunoenzyme method for the detection of the causative agent of tularemia]. / Ispol'zovanie immunofermentnogo metoda ELISA dlia vyiavleniia vozbuditelia tuliaremii.

The conditions of the enzyme-linked immunosorbent assay (ELISA) for the detection of Francisella tularensis were worked out. In the study of 27 strains differing in their biological characteristics, the sensitivity of the assay was determined, varying within the range of 1 X 10(4)--5 X 10(4) million cells/ml and exceeding the sensitivity of the currently used methods for the immunodiagnosis of tularemia by 1-2 orders. ELISA also proved to be a highly effective technique for the detection of the specific antigen in the organs of infected animals. The antigen was regularly detected in the organs of white mice, beginning from day 3 after their infection with the minimal doses of F. tularensis. The method may be recommended both for the identification of isolated cultures and for the early diagnosis of tularemia infection.

[Detection of the antigens of the causative agent of brucellosis by an immunoenzyme method]. / Vyiavlenie antigenov vozbuditelia brutselleza immunofermentnym metodom.

The conditions of the enzyme immunoassay for the detection of Brucella antigens have been selected, making it possible to detect these antigens both in solutions and in biological material within 3-4 hours. In guinea pigs infected with B. abortus 99 in a dose of 1,000 microbial cells, brucellar antigen has been detectable in the organs and blood serum of the animals as early as 24 hours after infection. This assay, if carried out under the optimal conditions, detects soluble brucellar polysaccharide antigen at a concentration of 1 ng/ml and Brucellae at a concentration of 5 X 10(4) microbial cells/ml in the presence of 200-fold surplus of other bacterial cells.

[Choice of the suitability criteria of polystyrene-based solid-phase carriers for performing immunoenzyme analyses]. / Vybor kriteriev prigodnosti tverdofaznykh nositelei na osnove polistirola dlia provedeniia immunofermentnogo analiza.

The authors discuss a tentative approach to the choice of criteria indicating the optimal suitability of different solid-phase carriers made of polystyrene for use in the enzyme immunoassay (EIA), viz. the dependence of specificity, sensitivity, reproducibility and reliability of EIA results on the adsorption properties, transparency expressed in percent and transparency variations of the plates under test. The evaluation of the carriers by four parameters is proposed with the use of assay plates manufactured by Nunc A/S (Denmark) for control. To ensure the objective evaluation of the suitability of polystyrene plates for use in EIA, the choice of uniform criteria is necessary.

[Immunoenzyme method in the diagnosis of human brucellosis]. / Immunofermentnyi metod v diagnostike brutselleza u liudei.

The optimum conditions for the determination of specific antibodies in the sera of brucellosis patients by means of enzyme immunoassay (EIA) have been selected. The comparative study of the specificity and sensitivity of EIA and other serological tests has demonstrated that EIA has high diagnostic effectiveness in the diagnosis of acute and chronic brucellosis. The presence of direct correlation between the results of EIA and Coombs' test is observed, which is indicative of the capacity of EIA for detecting both complete and incomplete specific antibodies. It should be pointed out that in all cases the titer of specific antibodies in EIA has been found to be 5-16 times higher than in Coombs' test, the passive hemagglutination test, and agglutination test.

[Development of an immunoenzyme method for detecting Francisella tularensis]. / Razrabotka immunofermentnogo metoda dlia vyiavleniia Francisella tularensis.

The conditions permitting the determination of F. tularesis cells by means of the enzyme immunoassay (EIA) in 3-5 hours have been established. Ways for enhancing the reliability of results obtained in the assay of the least possible amount of the test material have been proposed. The sensitivity and specificity of the rapid EIA technique permitting the determination of F. tularensis cells at a concentration of 20 000 cells/ml in the presence of other bacterial cells in 100-fold excess have been shown.

[Use of an immunoenzyme method on a solid-phase carrier for determining the tularemia antigen]. / Primenenie immunofermentnogo metoda na tverdofaznom nositele dlia opredeleniia tuliaremiinogo antigena.

The method of the highly sensitive (up to 10 ng/ml) and specific determination of soluble tularemia antigen, based on the use of "sandwich" type ELISA techniques, has been developed. The dependence of the specificity and sensitivity of the method on the degree of purification of antibodies and their peroxidase conjugates used in the assay has been studied. The study has revealed that the best results can be obtained with the use of purified IgG and its conjugate free of unbound peroxidase. Both foreign peroxidase preparations and type A enzyme manufactured in the USSR can be equally used as enzymatic labels.

[Determination of tularemia antibodies by an immunoenzyme method on a solid-phase carrier (ELISA)]. / Opredelenie tuliaremiinykh antitel immunofermentnym metodom na tverdofaznom nositele (ELISA).

ELISA "sandwich" techniques have been developed and the optimum assay conditions for detecting specific antibodies in human serum samples have been determined. The possibility of using these techniques for the determination of the level of antibodies to tularemia antigens in the sera of persons immunized with live tularemia vaccine has been shown. Statistically significant differences in the level of antibodies to tularemia antigen in the sera of immunized and nonimmunized persons have been established. The comparative study of five serological methods - ELISA, the agglutination test, the passive hemagglutination test, the immunofluorescence test and the defined antigen substrate sera ( DASS ) techniques - has revealed the advantage of ELISA, whose sensitivity has proved to be considerably higher than that of all other methods used in our work.

[Isolation and properties of polyphosphatase of Neurospora crassa]. / Vydelenie i nekotorye svoistva prolifosfatazy Neurospora crassa

Polyphosphatase (polyphosphate-phosphohydrolase) has been isolated from mycelium of Neurospora crassa and purified to homogenous state. The enzyme is shown to be strictly specific to high molecular weight inorganic polyphosphates. Km for phosphate in polymeric form is 6.8-10(-4) M. The molecular weight of this enzyme is 50 000 +/- 3000. To display its activity polyphosphatase requires the presence of bivalent cations of some metals, Mg2+ ions being the best activator with Co2+, Mn2+ and Fe2+ ions-slightly less effective.

[Possible physiological role of the "high molecular weight polyphosphate-polyphosphate phosphohydrolase" system in Neurospora crassa]. / O vozmozhnoi fiziologicheskoi roli sistemy "vysokomolekuliarnye polifosfaty-polifosfatfosforgidrolaza" u Neurospora crassa

The biosynthesis of polyphosphatase and the system of active transport of glucose is repressed in the cells of Neurospora crassa by glucose at a high concentration. There is a strict correlation between the activity of polyphosphatase and the initial rate of glucose active transport. Both systems are repressed during the growth of the mycelium on a glucose medium and are repaired on a medium without glucose. The latter process is inhibited by cycloheximide. Under various conditions of cultivation, the ratio between the activity of polyphosphatase and the initial rate of active transport of glucose remains close to unity. The experiments with 8-azaadenine have shown that the system of active transport of glucose does not require ATP. A possible physiological role of polyphosphatase in N. crassa is discussed.