Development and manufacture of erythrocyte pullorosis diagnosticum for the diagnosis of avian pullorosis typhoid

The article is devoted to the problems of diagnostics of poultry typhoid fever and the development of a technology for the manufacture of diagnosticum against this disease. A method of making erythrocyte diagnosticum for the diagnosis of poultry typhoid fever is shown, including obtaining the bacterial mass of Salmonella pullorum-gallinarum, isolating the antigenic fraction from it by treating the bacterial mass with a surfactant with the addition of soda or alkali in distilled water at 93-96 °C, followed by sensitization of formalinized erythrocytes, their purification and obtaining the target product in the form of 10% suspension, characterized in that 1-1.5% aqueous solution of Desmol is used as a surfactant to isolate the antigenic fraction, taken in a final weight concentration of 0, 1-0.3%, and the sensitization of formalinized erythrocytes is carried out in the presence of the sodium salt of chitosan succinate taken in a final weight concentration of 0.5-1.5%. In the industrial production of the diagnosticum at the initial stage, the bacterial suspension was necessarily mixed and the optical concentration was measured photometrically. The concentration of the surfactant solution was adjusted to 25 ml of microbial cells in 1 ml. At the second stage, antigen-sensitin was obtained and stored at 4 ° C. Sheep erythrocytes were used for sensitization. At the third stage, 20% formalized ram erythrocytes were obtained. Formalized erythrocytes were washed five times until the supernatant was completely cleared and sensitized. At the fourth stage, 300-500 ml was added to 1 liter of erythrocyte suspension for sensitization. sensitin and kept in a water bath at a temperature of 600С. At the fifth stage, the sensitized erythrocytes were washed to remove residual sensitin not associated with erythrocytes. Obtaining highly effective erythrocyte diagnostics for the diagnosis of avian pullorosis typhoid is an urgent problem. We have produced, improved and optimized the technology of industrial production of erythrocyte pullor antigen from the Salmonella pullorum-gallinarum strain for the diagnosis of avian pullorosis-typhus. We have theoretically substantiated and tested in production conditions a new method of resuspension, extraction, clarification of the bacterial mass. Used surfactants (surfactants) to obtain sensitin.

Selection of Specific Peptides for Coccidioides spp. Obtained from Antigenic Fractions through SDS-PAGE and Western Blot Methods by the Recognition of Sera from Patients with Coccidioidomycosis

Antigenic fractions of 100, 50, 37, and 28 kDa obtained through the SDS-PAGE method that were more frequently recognized by anti-Coccidioides antibodies in the sera of coccidioidomycosis patients were selected using western blotting. Subsequently, these bands were sequenced, and the obtained proteins were analysed by BLAST to choose peptides specific for Coccidioides spp. from among the shared aligned sequences of related fungi. A peptide specific for C. immitis was selected from the “GPI anchored serine-threonine rich protein OS C. immitis”, while from the “uncharacterized protein of C. immitis”, we selected a peptide for C. immitis and C. posadasii. These proteins arose from the 100 kDa antigenic fraction. From the protein “fatty acid amide hydrolase 1 of C. posadasii” that was identified from the 50 kDa antigenic fraction, a peptide was selected that recognized C. immitis and C. posadasii. In addition, the analysis of all the peptides (353) of each of the assembled proteins showed that only 35 had 100% identity with proteins of C. immitis and C. posadasii, one had 100% identity with only C. immitis, and one had 100% identity with only C. posadasii. These peptides can be used as diagnostic reagents, vaccines, and antifungals.

Structural and functional characterization of a novel scFv anti-HSP60 of Strongyloides sp.

Phage display is a powerful technology that selects specific proteins or peptides to a target. We have used Phage Display to select scFv (single-chain variable fragment) clones from a combinatorial library against total proteins of Strongyloides venezuelensis. After scFv characterization, further analysis demonstrated that this recombinant fragment of antibody was able to bind to an S. venezuelensis antigenic fraction of ~65 kDa, present in the body periphery and digestive system of infective larvae (L3), as demonstrated by immunofluorescence. Mass spectrometry results followed by bioinformatics analysis showed that this antigenic fraction was a heat shock protein 60 (HSP60) of Strongyloides sp. The selected scFv was applied in serodiagnosis by immune complexes detection in serum samples from individuals with strongyloidiasis using a sandwich enzyme-linked immunosorbent assay (ELISA), showing sensitivity of 97.5% (86.84–99.94), specificity of 98.81 (93.54–99.97), positive likelihood ratio of 81.60 and an area under the curve of 0.9993 (0.9973–1.000). Our study provided a novel monoclonal scFv antibody fragment which specifically bound to HSP60 of Strongyloides sp. and was applied in the development of an innovative serodiagnosis method for the human strongyloidiasis.

Pediatric Neurocysticercosis: Usefulness of Antibody Response in Cysticidal Treatment Follow-Up

Serum and urine samples were collected from 33 NCC patients before the albendazole treatment, 3–6 and 12 months PT. At 3 months PT, 24 (72.7%) patients had no detectable CT/MRI lesions and 9 (27.2%) patients had persistent lesions. Antibody response to crude soluble extract (CSE), excretory secretory (ES), and lower molecular mass (LMM) (10–30 KDa) antigenic fraction ofT. soliumcysticerci was detected in serum and urine samples by ELISA. Before the treatment, out of 33 NCC children, 14 (42.4%), 22 (66.6%), and 11 (33.3%) serum samples were found positive with the use of CSE, ES, and LMM antigen, respectively. At 3–6 months PT, positivity rate was 5 (15.1%), 2 (6%), and 4 (12.1%) and at 12 months PT, positivity rate was 5 (15.1%), 0, and 3 (9%) with the use of CSE, ES, and LMM antigen, respectively. There was no significant difference in the positivity with the use of three antigens in pretreatment and PT urine samples. The study suggests that the use of ES antigen to detect antibody in serum samples may serve better purpose to evaluate the therapeutic response in patients with NCC.

The determination of the degree of serological relationship among four carlaviruses using an immunoelectron microscope-decoration technique and cross-absorption tests

A very close serological relationship was found between CVB and PVS, while only a distant relationship was shown between CVB and CLV. Each of these viruses had its own characteristic major "antigenic fraction", distinguishing it from the others, and small "antigenic fraction" common to all four. The antiserum to CLV was shown to be the richest in various antigenic fractions, antisera against PVM and PVS had the same number of fractions but of different specificity and CVB-antiserum had the relatively smallest number of fractions.