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A simple method for stabilization of lyophilization or freezing/thawing sensitive enzyme-antibody conjugates is described. The method involves soaking of these reagents directly onto the placebo homoepathic sugar tablets followed by a brief air drying. This solid dry formulation is stable at elevated room temperature and is very convenient for storing or shipping small aliquots of these lyophilzation sensitive reagents.
The aim of these studies was to determine the performances of five commercial ELISA kits in the detection of BLV antibodies in serum samples. Different variants of ELI SA methods (indirect, blocking and double-well) were employed for these kits. Comparative investigations using 428 serum samples showed 418 consistent results (97.7%). Ten results (2.3%) proved to be inconsistent including 7 (1.6%) results which were totally different. The cause of these divergences in serological tests are discussed.
Congenitally abnormal fibrinogens with impaired fibrin monomer polymerization have been described to contain single amino-acid substitutions localized in certain positions of the γ275-330peptide region. To evaluate the role of the amino-acid sequence in the vicinity of Arg 275 in fibrin monomer polymerization, the peptide fragment corresponding to γ268-282was synthesized and used to obtain peptide-specific antibodies. These antibodies, when purified immunochemically on the immobilized peptide, bound to the intact fibrinogen and fibrin monomers with the same binding affinity. However, they did not recognize the Y268-282epitopes on the denatured and reduced fibrinogen molecules. The lack of influence of antipeptide antibodies on fibrin monomer polymerization indicates that the γ268-282peptide is not directly involved in the structure of the polymerization site in the D domain of fibrinogen. It is suggested that substitution of Arg275either by His or Cys in abnormal fibrinogens results probably in conformational changes which disturb a proper orientation of the polymerization site and reduce its expression.
The postulated intramolecular signaling in immunoglobulins generated by antigen binding has been controversial for years. The high heterogeneity of immune complexes as signaling systems and the requirement of the immobilized antigen form for efficient triggering of effector activity is likely the reason for the lack of clarity. Here we present new evidence supporting the notion of intramolecular signaling, based on the use of supramolecular dyes that bind to signal-derived specific sites in immunoglobulins.
Background. The strongest immunogen of the Rh system is the D antigen. It is found in several variants and categories, which makes it difficult to determine the correct RhD (Rh+) or RhD negative (Rh-) phenotype. Although only some of the varieties and types of this antigen are of clinical significance, it is important to determine the normal Rh phenotype in recipients and donors. The aim of this study was to determine the frequency of weak D antigen in a population of potential recipients. Material and methods. The study group consisted of selected blood recipients in whom weak expression of the D antigen or its antibody was detected. In order to estimate the expression of antigen D, the blood was analyzed in the laboratory of the Regional Center of Blood Donation and Blood Treatment in Lublin. Blood from 220 potential recipients (149 women and 71 men) were used in the conducted research. The clinical material from the Laboratory of Transfusion Serology at the Provincial Specialist Hospital in Biała Podlaska was also used. Results. The presence of a weak D was confirmed in 21 recipients. 4 cases of weak D were confirmed among recipients of blood transplant, while 17 cases among those who did not have blood transfusions. There were significant differences in the occurrence of the weak D in relation to the transfusion in both women (χ² = 18.34 df = 2, p = 0.0001) and men (χ² = 17.25) . Conclusions. The correct determination of the RhD+ or RhD- phenotype is important for pregnant women who should be subjected to immunoprophylaxis of maternal-fetal conflict when a weak D is detected. In order to avoid post-transfusion complications among recipients, it is necessary to choose serologically and phenotypically crossed-matched blood components.
Background: Lyme disease is a multi-organ disease caused by spirochetes, Borrelia burgdorferi sensu lato, transmitted by Ixodes, with its clinical picture including involvement of the skin, joints, nervous system and heart. Laboratory diagnostic tests for Lyme disease are mainly based on the detection of anti-Borrelia burgdorferi antibodies by means of serological methods. Aim of the work: assessment of the level of antibodies against specific B. burgdorferi s.l. antigens in persons with suspected Lyme disease. Material and methods: the tested group consisted of 98 patients with suspected Lyme disease. During the first phase of the tests, anti-Borrelia burgdorferi IgM/IgG antibodies were marked using ELISA method, and positive and uncertain results were confirmed by Westernblot test (Wb). Results: anti-B. burgdorferi IgM/IgG antibodies were present in 60 patients (61.2%). IgM and IgG antibodies were detected as positive in 8 (8.1%) and 35 (35.7%) patients respectively. IgM and IgG were co-present in 6 persons (6.1%), including 2 persons (2%) with positive results in both classes. All patients with positive IgM (12 persons) had anti-OspC antibodies, and 2 patients had, in addition, anti-p31 antibodies. In patients with positive IgG the results were as follows: antibodies against antigen p17 - 77% of cases, VlsE - 74%, p30 - 46%, p39 - 44%, p83 - 38%, p19 - 31%, OspC/p25- 28%, p31 - 23%, p21 - 8%. Conclusions: laboratory diagnostic tests for Lyme disease must be performed in accordance with the current standards, positive and uncertain results must be confirmed by Westernblot test. Results of lab tests must correlate with patient’s symptoms.
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