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The protein antigens of BLV were detected in the lysates of bovine lymphocytes stimulated in vitro using the Western blot method. Specific antibodies against BLV were detected in the sera of animals by the ELISA test. Protein bands of the molecular weight 72, 51, 35 and 24 kD were detected by the rabbit antiserum against BLV in the blots from infected lymphocytes and FLK cells. Thе proteins mentioned above were not detected in the lysates of lymphocytes of uninfected animals. Densitometry analysis of immunoblots from FLK cells indicated that a minimal amount of 100 ug of gp51 viral protein can be discovered. The presence of BLV antigens and specific antibodies were detected in 18 of 25 animals. The antibodies were also found in four cows of the remaining seven animals whose lymphocytes were free from BLV proteins.
The aim of the study was to evaluate the application of SDS-PAGE electrophoresis and western-blotting for serological diagnostics of glanders. Six commercial Burkholderia mallei antigens, eight anti-B. mallei sera and two negative sera were used in the study. In the first part of the study electrophoretic profiles of antigens in SDS-PAGE were determined at a gradient of 4% to 20% of polyacrylamide gel. Optimal conditions of western-blotting reaction were determined. In the case of all examined antigens and anti-B. mallei sera, the presence of protein fractions in the range from 42 to 33 kDa and one band of the molecular weight of 16 kDa were demonstrated by immunoblotting reaction. Additional bands of the different molecular weight were also observed. The antigens examined did not show reaction with negative sera. In the next part of the study, a sensitivity of the western-blotting method and the CFT were compared. The western-blotting method was two - four times more sensitive than the CFT.
Biotinylated surface and somatic protein extracts of the nematode Capillaria resecta were analysed by SDS-PAGE followed by Western blotting in order to examine their antigenic character. The antigens were probing with serum of C. resecta naturally infected birds (jackdaws). Surface and somatic antigens were recognised by serum IgG, IgA and IgM antibodies. The most intense reactivity of sera was evident with TBS-soluble surface proteins. The present paper has shown that surface proteins of C. resecta are more immunogenic than somatic and they are able to elicit stronger immune response in the bird's body.
The beginning of bee virology dates back to 1963, when the chronic bee paralysis virus (CBPV) was discovered. To date about 15 viruses of honey bee Apis mellifera have been identified. Those viruses persist in bee populations usually as unapparent infections, but under some conditions, e.g. severe Varroa infestation, they can cause adult bee and brood mortality. Some of the viruses can occur on their own, others together with another pathogen. Almost all bee viruses contain positive stranded RNA, only filamentous virus has DNA. On the basis of recent studies some of these viruses were classified in the newly created family Dicistroviridae, which includes the single genus Cripavirus, and in the floating genus Iflavirus unassigned to any family. However, most bee viruses are not classified yet. Agar gell immunodiffusion test (AGID) is a very convenient method for the diagnosis of bee virus infections. This technique is easy to use but has low sensitivity and requires the production of specific antisera. In research into bee viruses other techniques are also employed, e.g. ELISA, western blotting, and RT-PCR.
Regulacje prawne narzucające obowiązek znakowania produktów żywnościowych zawierających GMO przyczyniły się do rozwoju rzetelnych i czułych technik jego detekcji. W artykule opisano immunodiagnostyczne metody idealne do jakościowej i ilościowej analizy GM żywności, takie jak ELISA, LFD, Western Blot. Wskazano ich zalety i wady oraz limity detekcji uzyskane dla produktów żywnościowych badanych z ich udziałem. Opisano także alternatywne podejścia badawcze, takie jak NIRS, skuteczne w detekcji nieznanej dotychczas genetycznej modyfikacji żywności. W artykule porównano również mocne i słabe strony metod immunodiagnostycznych oraz opartych na analizie DNA.
The article concentrates on diagnostic aspects of the major mycoplasma diseases in ruminants. Among them particular attention is given to: bovine contagious pleuropneumonia and contagious agalactia of sheep and goats, and an involvement of mycoplasmal etiological agent in bovine respiratory disease (BRD). The diagnosis of mycoplasma infections is based on serological methods such as: Elisa test, complement fixation test (CFT), and Western blot. The culture and identification of Mycoplasma spp. are possible through the use of specific liquid or solid Eaton’s media. The doubtful cases are confirmed by molecular biology methods (PCR and DGGE).
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