Search results for "Dot blot"
showing 10 items of 11 documents
Development of an Analytical Assay for Electrochemical Detection and Quantification of Protein-Bound 3-Nitrotyrosine in Biological Samples and Compar…
2020
Reactive oxygen and nitrogen species (RONS) cause oxidative damage, which is associated with endothelial dysfunction and cardiovascular disease, but may also contribute to redox signaling. Therefore, their precise detection is important for the evaluation of disease mechanisms. Here, we compared three different methods for the detection of 3-nitrotyrosine (3-NT), a marker of nitro-oxidative stress, in biological samples. Nitrated proteins were generated by incubation with peroxynitrite or 3-morpholino sydnonimine (Sin-1) and subjected to total hydrolysis using pronase, a mixture of different proteases. The 3-NT was then separated by high performance liquid chromatography (HPLC) and quantifi…
Meat allergy associated with galactosyl‐α‐(1,3)‐galactose (α‐Gal)—Closing diagnostic gaps by anti‐α‐Gal IgE immune profiling
2017
Background Glycoproteins and glycolipids of some mammalian species contain the disaccharide galactosyl-α-(1,3)-galactose (α-Gal). It is known that α-Gal is immunogenic in humans and causes glycan-specific IgG and also IgE responses with clinical relevance. α-Gal is part of the IgE-reactive monoclonal therapeutic antibody cetuximab (CTX) and is associated with delayed anaphylaxis to red meat. In this study, different α-Gal-containing analytes are examined in singleplex and multiplex assays to resolve individual sensitization patterns with IgE against α-Gal. Methods Three serum groups, α-Gal-associated meat allergy (MA) patients, idiopathic anaphylaxis (IA) patients with suspected MA, and non…
Lack of hepatitis B virus DNA sequences in sera from patients with acute and chronic liver diseases diagnosed as non-A, non-B-hepatitis
2008
— The sera of 15 individuals with transfusion-associated acute or chronic non A, non B hepatitis, which lacked hepatitis B virus markers, were tested for hepatitis B virus DNA by dot blot hybridization test. Three sera of two patients positive in this test, however, also gave positive results when the labeled plasmid was used as probe instead of labeled HBV-DNA, indicating false positive results in the initial test. In conclusion, the data indicated that sera of patients with confirmed non A, non B hepatitis do not contain DNA-sequences in the serum hybridizing with HBV-DNA.
DNA-DNA hybridization and ribotyping of Acidovorax delafieldii isolates from eels and aquatic environments
1992
Eleven strains of Acidovorax isolated from a freshwater lake and from healthy and diseased European eels in a hatchery were investigated by genotypic methods. DNA-DNA slot blot hybridization confirmed the identity of all strains as A. delafieldii. Isolates were heterogeneous in EcoR1 ribopatterns with four types recognizable. Most freshwater and healthy eel strains had the same ribotype but differed from the disease-associated strains. Typical ribopatterns of A. delafieldii contained 1·3, 1·5, and 3·2 kb fragments which provided useful additional taxonomic markers for species identification.
Sex Determination by Genomic Dot Blot Hybridization and HLA DQα Typing by PCR from Fixed Tissues
1992
Recent advances in molecular biology methods have significantly increased the ability to detect genetic variation at the genomic level for forensic purposes. However, the quality requirements for blood, fresh or frozen tissue as a source of DNA are a practical limitation for typing the victim in order to conduct investigations on unsolved cases. Since paraffin embedded specimens are easily obtainable the ability to study this material would be of great value in current forensic practice.
Klinische Bedeutung des Hepatitis-B-Virus-DNS-Nachweises im Serum von Kindern mit chronischer Hepatitis B
1992
206 sera from 172 children with chronic hepatitis B infection were tested for HBV DNA by dot blot hybridization. 111 were positive and 95 negative for HBV DNA. 103 (78.6%) of the positive patients had HBeAg and 5 (7.7%) anti-HBe. In 60 (92.3%) of the anti-HBe positive sera no HBV DNA could be detected. Children with elevated liver enzymes had HBV DNA in 80.1%, whereas in 71.6% of the chronic HBsAg carriers with normal liver enzymes no HBV DNA was found. In 87 of the 95 dot blot negative patients polymerase chain reaction was performed. 73 (83.9%) children of this group were HBV DNA positive. All HBeAg positive patients and those with elevated aminotransferases had HBV DNA in their serum. 56…
CHARACTERIZATION OF CHRONIC HEPATITIS B IN CHILDHOOD USING MOLECULAR BIOLOGY TECHNIQUES
1992
The introduction of molecular biology techniques in the diagnostics of chronic hepatitis B virus infection proved HBV DNA to be the most sensitive marker of viral replication and infectivity. The aim of our study was to characterize the HBV DNA status in children with chronic hepatitis B with various molecular biology techniques in relation to conventional HBV markers. Methods: 206 sera of 172 and liver tissue of 108 children with chronic hepatitis B infection were investigated by dot blot-, Southern blot-, and in situ hybridization. In dot blot and Southern blot negative specimens polymerase chain reaction (PCR) was performed. Results: 111 of the 206 sera were positive for HBV DNA by dot b…
A methodological strategy for PAH genotyping in populations with a marked molecular heterogeneity of hyperphenylalaninemia.
2001
Abstract The elucidation of the molecular basis of hyperphenylalaninemia in various world populations (PKU Consortium Database: http://www.mcgill.ca/pahdb/) has revealed a remarkable molecular heterogeneity at the locus encoding for phenylalanine hydroxylase. As a consequence, genotyping of HPA patients has prompted the establishment of an impressive number of mutation detection protocols. In spite of the large variety of methods proposed so far, no comprehensive strategy has been yet developed for the detection of PAH gene mutations. Therefore, new approaches, combining the advantages of individual methods are required, especially in populations with a high number of PAH gene mutations. In…
The kinetics of antibody production in mucus and serum of European eel (Anguilla anguilla L.) after vaccination against Vibrio vulnificus: developmen…
2003
Abstract Vibrio vulnificus serovar E, a bacterial pathogen for eels cultured in intensive systems, is transmitted through water and enters into new hosts mainly via gills. The main objective of this work was to study the kinetics of antibody production to V. vulnificus in serum and mucus and their relationship with protection after vaccination. To quantify local mucus antibodies, a new ‘in situ’ dot blot immunoassay using image analysis has been developed. This assay was applied to measure antibody production in the skin zone next to the gills. We found that (i) the immune response in mucus was faster (peak at days 3–4) and shorter in duration (titres significantly elevated up to day 5 and …
Application of DNA techniques for identification using human dental pulp as a source of DNA
1992
Dental pulp tissue could be obtained in most cases from materials obtained under experimental conditions and from forensic casework (air accidents, burned and putrefied bodies). Teeth extracted during dental treatment (n = 30) were stored for 6 weeks and 4 years at room temperature. In addition teeth (n = 10) extracted from jaw fragments that had been stored for 15 years at room temperature, and teeth extracted post mortem from actual identification cases (n = 8) were investigated. Following extraction from dental pulp tissue the DNA concentration was measured by fluorometry. The amount of DNA obtained from the dental pulp tissue of a single tooth varied from 6 micrograms to 50 micrograms D…