[Comparative assessment of DNA extraction methods for identification of glanders and melioidosis etiological agents by PCR].

Pathogenic Burkholderia are considered as a cause of dangerous infections and potential agents of bioterrorism. Comparative assessment of different methods of extraction and purification of DNA for PCR analysis of pure cultures and samples contaminated by etiological agents of glanders and melioidosis was performed. Samples of soil and food artificially contaminated by pathogenic Burkholderia as well as organs of infected animals were tested. DNA was extracted by methods of boiling, nucleosorption with presence of guanidine thiocyanate, guanidine thiocyanatephenol extraction, guanidine thiocyanate-phenol extraction with additional purification of DNA by nucleosorption. Amplification was performed by "Flash" technique and detector of fluorescence was used for analysis of PCR products. Utilization of the recommended methods of preparation depending on the nature of sample let to detect by the "Flash" technique the etiological agents of glanders and melioidosis in concentration =10(3) microbial cells per ml. Choice of DNA extraction and purification methods is determined by type of a sample and presence in it of admixtures inhibiting PCR.

[The properties of the extracellular alkaline phosphatase of the causative agent of melioidosis]. / Izuchenie svoistv vnekletochnoi shchelochnoi fosfatazy vozbuditelia melioidoza.

After growing P. pseudomallei VPA on solid medium extracellular alkaline phosphatase with a molecular weight of 93,000 AMU was isolated, and practically purified from the extract of this medium by precipitation with ammonium sulfate, subsequent gel chromatography and concentration on membrane filters. The optimum conditions for enzymatic reaction were found to be pH 9.0 and a temperature of 50 degrees C. The enzyme was resistant to freezing and to heating at a temperature of up 60 degrees C for 30 minutes, as well as to the action of pH 3.0-10.5, but became completely inactivated after heating at 90 degrees C for 10 minutes and incubation at pH 2.0 for 20 hours.

[Immunofluorescence method of detecting cholera enterotoxin]. / Immunofliuorestsentnyi metod obnaruzheniia kholernogo énterotoksina.

The quantitative immunofluorescent assay for the determination of cholera enterotoxin is proposed. The assay is based on the selective sorption of cholera enterotoxin by gangliosides incorporated into polyacrylamide granules. The preliminary treatment of gangliosides with neuraminidase enhances the sensitivity of this assay. The assay permits the detection of cholerigen in an amount of 20 ng.

[Use of neuraminidase for improving the properties of the erythrocytic ganglioside diagnosticum]. / Ispol'zovanie neiraminidazy dlia uluchsheniia svoistv eritrotsitarnogo gangliozidnogo diagnostikuma.

In this work the possibility of using neuraminidase for increasing the content of ganglioside GM1 in the mixture of gangliosides used for the sensitization of erythrocytes has been studied. The study has revealed that the treatment of gangliosides with neuraminidase is sufficient for obtaining active hemosensitin; there is no need for the purification of the preparation by gel filtration.

[Peptidase activity of Coccidioides immitis]. / Peptidaznaia aktivnost' Coccidioides immitis.

Glycyl-L-leucinehydrolase consisting of three molecular units was extracted from C. immitis solid cultural medium. During fractionation in polyacrylamide gel of the enzyme-containing extract a 50-fold purification of the enzyme isoform with molecular weight 12,800 is achieved. The enzyme is heat-stable, active in the narrow pH range and hydrolizes peptide bonds containing glycine. Its activity is not inhibited by none of the protease inhibitors tested.