RESUMEN
The efficacy of a kit of Opisthorchiasis-CIC-EIA-Best reagents was evaluated using 270 sera from patients in the study and control groups. The kit showed a sufficient sensitivity (not less than 87.2%) and a high specificity (not less than 97.9%). The use of the above kit of the reagents for enzyme immunoassay in practical healthcare enables one to increase detection rates among the infested subjects on comprehensive examination of those with suspected opisthorchiasis.
Asunto(s)
Anticuerpos Antihelmínticos/inmunología , Complejo Antígeno-Anticuerpo , Antígenos Helmínticos/inmunología , Técnicas para Inmunoenzimas , Opistorquiasis , Opisthorchis/crecimiento & desarrollo , Animales , Complejo Antígeno-Anticuerpo/inmunología , Estudios de Casos y Controles , Heces/parasitología , Humanos , Opistorquiasis/sangre , Opistorquiasis/diagnóstico , Opistorquiasis/inmunología , Opistorquiasis/parasitología , Recuento de Huevos de Parásitos , Federación de Rusia , Sensibilidad y EspecificidadAsunto(s)
Opistorquiasis/epidemiología , Opisthorchis/aislamiento & purificación , Adolescente , Adulto , Animales , Anticuerpos Antihelmínticos/sangre , Pueblo Asiatico , Niño , Humanos , Persona de Mediana Edad , Opistorquiasis/sangre , Opistorquiasis/etnología , Opisthorchis/inmunología , Estudios Seroepidemiológicos , Población BlancaRESUMEN
Sensitizing and virus-neutralizing properties of IgG isolated from the sera of goats immunized with Ebola virus and a relevant gammaglobulin prepared by ethanol fractionation were compared. The ratio of the virus-neutralizing activities of subclasses IgG2, IgG1a, and IgG1b was 100:10:1. Anaphylactogenic activity of IgG2 in the immediate type hypersensitivity test in guinea pigs was 2-fold lower than that of IgG1a and IgG1b. Goat gammaglobulin to Ebola virus, consisting from IgG2 antibodies by more than 50%, possessed virus-neutralizing and sensitizing characteristics compatible to IgG2.
Asunto(s)
Anticuerpos Antivirales/inmunología , Ebolavirus/inmunología , Cabras/inmunología , Inmunoglobulina G/inmunología , Anafilaxia , Animales , Anticuerpos Antivirales/administración & dosificación , Anticuerpos Antivirales/biosíntesis , Cabras/sangre , Cobayas , Inmunización , Inmunoglobulina G/administración & dosificación , Inmunoglobulina G/biosíntesis , Pruebas de NeutralizaciónRESUMEN
Homogeneous (according to PAGE) capsid and surface viral proteins were isolated from concentrated purified suspensions of tick-borne encephalitis and Venezuelan equine encephalomyelitis viruses by one-stage reversed-phase HPLC. The amino acid composition and the sequences of their N-terminal parts were determined.
Asunto(s)
Virus de la Encefalitis Equina Venezolana/química , Virus de la Encefalitis Transmitidos por Garrapatas/química , Proteínas Estructurales Virales/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Proteínas Estructurales Virales/químicaRESUMEN
Whether purified antigens of Lamblia intestinalis trophozoites can be used to detect these antibodies by immunoassay. The drugs of immunodominant Lamblia antigens were prepared by anion-exchange chromatography of solubilized trophozoite components and they are mainly presented by proteins having molecular weights of 70, 56, and 49 kD. Immunoassay using these antigens revealed antibodies to Lamblia trophozoite antigens in sera of 87.6% of patients with lambliasis (its diagnosis was established on the basis of microscopic data on the duodenal content) and only in 16.2% of clinically healthy blood donors. Twenty six sera from patients with trichomoniasis having high levels of antibodies to trichomonad antigens were studied to evaluate the specificity of this method for detection of antibodies. It has been found that the proportion of subjects in this group who have also antibodies to Lamblia antigens does not greatly differ from that of healthy blood donors (19.2 and 16.2, respectively).
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/inmunología , Giardia lamblia/inmunología , Estadios del Ciclo de Vida/inmunología , Animales , Animales Lactantes , Antígenos de Protozoos/aislamiento & purificación , Donantes de Sangre , Epítopos , Heces/parasitología , Giardia lamblia/crecimiento & desarrollo , Giardia lamblia/aislamiento & purificación , Giardiasis/diagnóstico , Humanos , Técnicas para Inmunoenzimas , Ratones , Tricomoniasis/diagnósticoRESUMEN
The authors developed a method for obtaining highly specific polyclonal antibodies reacting with different sites of human chorionic gonadotropin (HCG) and an immunometrical method for measuring HCG in human biological fluids, based on the use of these antibodies. The sensitivity of the method is 10-15 IU/liter HCG, specificity 100%, no cross reactions with LH or FSH were observed. The method was tried in testing urine samples for HCG in women at 1-2 to 34 weeks of gestations and in one cancer patient with the diagnosis of vesical mole.
Asunto(s)
Gonadotropina Coriónica/orina , Animales , Anticuerpos Monoclonales , Cromatografía de Afinidad , Femenino , Humanos , Sueros Inmunes/inmunología , Técnicas para Inmunoenzimas , Embarazo , Conejos , Sensibilidad y EspecificidadRESUMEN
Seroprevalence to Toxocara antigens was studied among Novosibirsk inhabitants and a focus of toxocariasis was found in the southern regions of west Siberia. Analyzing the sera of opisthorchiasis patients revealed antibodies to Toxocara antigens, with high titers (1:800 or more) in 1.80% of cases. Immunoblotting of these sera and those from patients with toxocariasis and opisthorchiasis, which were seropositive in one of the above helminths showed that there were mixed human infections with Toxocara and Opisthorchis in this area.
Asunto(s)
Reservorios de Enfermedades , Larva Migrans Visceral/epidemiología , Opistorquiasis/epidemiología , Animales , Anticuerpos Antihelmínticos/sangre , Eosinofilia/epidemiología , Eosinofilia/inmunología , Humanos , Larva Migrans Visceral/inmunología , Opistorquiasis/inmunología , Opisthorchis/inmunología , Prevalencia , Estudios Seroepidemiológicos , Siberia/epidemiología , Toxocara canis/inmunología , Población Urbana/estadística & datos numéricosRESUMEN
Enzyme immunoassay (EIA) test systems for the detection of antigens of and antibodies to Ebola virus were developed and tried. The test system for the detection of Ebola virus antigens based on direct solid-phase EIA detects viral antigens in culture fluid of infected Vero cells, in the blood sera, and in homogenates of infected tissues. Use of this test system allows detection of at least 10 ng of viral proteins or 5.0 x 10(3) to 1.0 x 10(4) PFU/ml in infectious material. The test system is prepared on the basis of protein A - horseradish peroxidase conjugate. It is universal for the testing of animal and human sera and is characterized by high resolution and reproducibility of results. It allows detection of antibodies to Ebola virus starting from days 8-9 of infection. A higher sensitivity of direct solid-phase EIA in comparison with complement fixation or indirect immunofluorescence tests is demonstrated.
Asunto(s)
Ebolavirus/inmunología , Fiebres Hemorrágicas Virales/diagnóstico , Animales , Anticuerpos Antivirales/análisis , Antígenos Virales/análisis , Línea Celular , Chlorocebus aethiops , Fiebres Hemorrágicas Virales/virología , Humanos , Técnicas para Inmunoenzimas , Reproducibilidad de los Resultados , Células VeroRESUMEN
Serum samples containing Ebola Virus neutralizing antibodies were prepared by prolonged immunization of goats with 10% liver homogenate from guinea pigs infected with Ebola virus. Differences in IgG fractions of normal and hyperimmune caprine blood sera were detected. Analytical chromatography on Polysil SA and immunodiffusion showed the presence of three IgG-containing fractions in hyperimmune sera. Immunochemical properties of these fractions were studied by solid-phase enzyme immunoassay and neutralization test. Antibodies to viral antigens were referred to IgG2 and IgG1a, and virus neutralizing properties were found mainly in IgG2 antibodies. Immunoglobulins were isolated from hyperimmune serum by alcohol sedimentation. The principal IgG fraction was found to contain much lower levels of antibodies to guinea pig liver antigens than intact serum, but at the same time it was characterized by a higher neutralization index.
Asunto(s)
Ebolavirus/inmunología , Inmunoglobulinas/metabolismo , Animales , Antígenos Virales/inmunología , Cabras , Cobayas , Sueros Inmunes , Inmunización , Inmunoquímica , Técnicas para Inmunoenzimas , Inmunoglobulinas/inmunología , Pruebas de NeutralizaciónRESUMEN
During the experiments 4 murine and 3 rat hybridomas producing monoclonal antibodies (MAb) against the protein p24 of human immunodeficiency virus type 1 (HIV-1) have been obtained. Using the immunoblotting technique, it was established that all the species of MAb reacted with the same viral proteins which are derivatives of gag gene--p24 and p55. The properties of MAb have been studied in competitive binding. Their ability of binding to different fragments of the gag protein produced by the recombinant plasmids in E. coli cells have been investigated in ELISA. The analysis of the findings suggests that the HIV-1 protein p24 contains at least 3 antigenic epitopes. All species of MAb reacted with 3 different HIV-1 strains and 2 HIV-1 isolates, but failed with 2 different HIV-2 strains. The only MAb NS5E4 can be used as an immunosorbent in the antigenic capture reaction.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/inmunología , Anticuerpos Monoclonales/inmunología , Proteína p24 del Núcleo del VIH/análisis , VIH-1/inmunología , Síndrome de Inmunodeficiencia Adquirida/microbiología , Animales , Anticuerpos Monoclonales/biosíntesis , Unión Competitiva/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Epítopos/análisis , Epítopos/inmunología , Proteína p24 del Núcleo del VIH/inmunología , Humanos , Hibridomas/inmunología , Immunoblotting/métodos , Ratones , Ratones Endogámicos BALB C , Ratas , Ratas EndogámicasAsunto(s)
Escherichia coli/metabolismo , Genes Sintéticos , Interferón-alfa/genética , Biosíntesis de Péptidos , Secuencia de Aminoácidos , Aminoácidos/análisis , Secuencia de Bases , Cromatografía Líquida de Alta Presión , Expresión Génica , Humanos , Hidrólisis , Datos de Secuencia Molecular , Péptidos/metabolismo , Plásmidos , Homología de Secuencia de Ácido NucleicoRESUMEN
Constituent proteins of human influenza virus A have been separated by reverse-phase HPLC on Polysil ODS-500. Their homogeneity is confirmed by the data of amino acid composition, Edman analysis and gel electrophoresis.
Asunto(s)
Virus de la Influenza A/metabolismo , Proteínas Virales/aislamiento & purificación , Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Electroforesis en Gel de Poliacrilamida , Datos de Secuencia Molecular , Albúmina Sérica Bovina/aislamiento & purificación , Proteínas Virales/genéticaRESUMEN
Three peptides corresponding to the sequences 124-144, 124-138, 129-144 of the human leukocyte interferon alpha 2 (IFN-alpha 2) were synthesized. The synthesis was performed by DCC-HOBT coupling of protected peptide segments in solution. The segments were obtained by the active ester coupling methodology using base-labile 2-[4-(phenylazobenzyl)sulfonyl]ethyl (Pse) group as carboxyterminal protection. After complete deprotection with 1 M methanesulphonic acid in trifluoroacetic acid--thioanisol--m-cresol mixture the peptides were purified by reversed-phase chromatography. The studies of interaction of the peptides with rabbit antiserum against IFN-alpha 2 revealed at least one minor antigenic determinant within the 124-144 region of IFN-alpha 2 amino acid sequence. Rabbit antisera developed against peptides 124-138 and 129-144 showed ability of binding recombinant IFN-alpha 2 and neutralizing its antiviral activity. Free peptides or their conjugates with bovine serum albumine did not display antiviral activity, neither could they inhibit the activity of IFN-alpha 2.
Asunto(s)
Interferón-alfa/síntesis química , Fragmentos de Péptidos/síntesis química , Secuencia de Aminoácidos , Antivirales , Unión Competitiva , Epítopos/química , Epítopos/inmunología , Inmunoquímica , Interferón alfa-2 , Interferón-alfa/química , Interferón-alfa/inmunología , Interferón-alfa/farmacología , Datos de Secuencia Molecular , Fragmentos de Péptidos/química , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/farmacología , Proteínas RecombinantesRESUMEN
M13B1 vector based on the filamentous phage M13 has been constructed. M13B1 phage carries the gene of resistance to ampicillin and contains the unique site of recognition for BamHI restriction endonuclease in gene VIII coding for the major coat protein. BamHI restriction site has been inserted into the gene of the major coat protein by means of oligonucleotide directed mutagenesis. The synthetic DNA fragment coding for the model peptides has been inserted through BamHI site into the M13B1 DNA. The possibility of inserting foreign peptides into the N-terminus at maintaining the viability of hybrid phages has been shown. The differences in specificity of the recombinant phage maturation have been determined by analysing the amino acid sequence of B-protein.
Asunto(s)
Proteínas de la Cápside , Cápside/genética , Colifagos/genética , ADN Viral/genética , Proteínas de la Membrana/genética , Ingeniería de Proteínas , Secuencia de Aminoácidos , Secuencia de Bases , Southern Blotting , Datos de Secuencia Molecular , Mutación , Recombinación GenéticaRESUMEN
The antigenic properties of the synthetic peptides corresponding to the regions 122-133, 136-147, 154-164 of the virus A/Aichi/2/68 hemagglutinin heavy chain have been studied. The 122-133 and 136-147 peptides comprise together almost whole antigenic determinant A, while the 154-164 peptide is a part of determinant B. Rabbits immunized by the peptides conjugated with carrier-protein BSA gave the high immune response to hapten-peptides. Each antiserum reacted only with homologous conjugate. All the antipeptide serums reacted with the virus A/Aichi/2/68 fixed on the base. Conjugate of the 136-147 peptide reacted with the rabbit antiserum against the virus A/Aichi/2/68 rendering the direct evidence to location of at least one hemagglutinin antigenic determinant in the region 136-147.
Asunto(s)
Antígenos Virales/inmunología , Epítopos/análisis , Hemaglutininas Virales/inmunología , Virus de la Influenza A/inmunología , Péptidos/inmunología , Animales , Embrión de Pollo , Humanos , Péptidos/síntesis químicaRESUMEN
Solution syntheses of [Leu]enkephalin and its [D-Ala2]analogue were accomplished using a new 2-(4-chlorophenyl)sulfonylethoxycarbonyl (Cps) base-labile group for amino protection and a chromogenic acid-labile 4-(4-phenylazo)benzyloxybenzyl (Abz) group for carboxyl protection. The syntheses were performed by stepwise attachment of Cps-amino acids, which were introduced as pentachlorophenyl esters or as dicyclohexylammonium salts in the presence of tris(dimethylamino)chlorophosphonium perchlorate. To remove Cps-group, Abz-esters of Cps-peptides were treated with two molar equivalents of 1,8-diazabicyclo[5.4.0]undec-7-ene in dimethylformamide followed by neutralization of the base with an excess of 1-hydroxybenzotriazole; the deblocked amino components were then used without isolation. Fully deblocked pentapeptides were purified and characterized by HPLC, FAB mass spectra and amino acid composition.
Asunto(s)
Leucina Encefalina-2-Alanina/análogos & derivados , Encefalina Leucina/análogos & derivados , Encefalina Leucina/síntesis química , Aminoácidos , Ácidos Carboxílicos , Fenómenos Químicos , Química , Cromatografía Líquida de Alta Presión , Indicadores y Reactivos , Espectrometría de MasasRESUMEN
Peptides corresponding to sequences 122-133, 136-147, and 154-164 of the heavy chain of hemagglutinin of the A/Aichi/2/68 (H3N2) influenza virus have been synthesized by stepwise elongation of the peptide chain with Boc-amino acid activated esters or by condensation of peptide blocks by DCC/HOBt-method. A coloured C-protecting group, 2-[4-(phenylazo)-benzylsulfonyl]ethyl (PSE), was used, which is convenient in purification of synthetic peptides. After removal of terminal N-and C-protecting groups the side-protecting residues were cleaved off with 1 M trifluoromethanesulfonic acid in trifluoroacetic acid containing 10% thioanisole. Crude products were purified by preparative reversed-phase liquid chromatography. Synthesized peptides were conjugated with BSA.