Search results for "Hybridization probe"
showing 10 items of 26 documents
PCR-Typing of the Human HLA-DQα Locus: Population Genetics and Application in Forensic Casework
1991
Multi- and single-locus probes recognizing highly polymorphic DNA sequences throughout the genome ([1–3]; C. Rittner, this volume) have become powerful tools for paternity testing and forensic stain analysis. In forensic casework, however, DNA probe technology can often not be applied, since genomic DNA extracted from stain material exposed to conditions of high temperatures and humidity is degraded. Also, the amount of DNA to be typed may not be sufficient to use DNA probe technology, e.g. if extracted from minute blood or sperm stains, from single hairs or from cell smears on microscope slides.
C4 DNA RFLP reference typing report.
1990
One hundred and three individual DNA samples (including 23 families) were studied at the gene level during the reference typing of the fourth component of human complement at the VIth Complement Genetics Workshop in Mainz (1989). All samples were analyzed with the restriction enzyme Taq I and with two DNA probes recognizing the 5' ends of both C4 genes and the two adjacent 21-hydroxylase genes. This RFLP is informative for the number of C4 genes as well as for their respective gene size. We found a high degree of variation regarding the number of C4 genes, i.e. haplotypes with 1-3 structural C4 genes of 16 or 22 kb size. By correlating these haplotypes to the complotypes obtained by protein…
Cytogenetic and molecular findings related to rhabdomyosarcoma. An analysis of seven cases.
2003
Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma in childhood. Histologically, it is subdivided histologically into two main subtypes: alveolar (ARMS) and embryonal (ERMS). ARMS is characterized by t(2;13)(q35;q14) or its variant t(1;13)(p36;q14), which fuse PAX3 and PAX7, respectively, with FKHR to produce chimeric genes. ERMS is frequently associated with loss of heterozygosity of 11p15.5. We investigated seven RMS (three ARMS and four ERMS) by means of cytogenetic, fluorescence in situ hybridization, and molecular analyses, including the study of the main genes implicated in the G1- to S-phase cell cycle transition, and correlated these studies with pathologic findings and c…
Molecular analysis in patients with mucopolysaccharidosis type II suggests that DXS466 maps within the Hunter gene
1993
Hunter disease is an X-linked mucopolysaccharidosis caused by deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS). Using the IDS cDNA and DNA probes corresponding to loci flanking the IDS locus, we performed molecular genetic studies in two patients with Hunter syndrome. An interstitial deletion spanning the middle part of the IDS gene was found in the first patient. The second patient carries a gross gene rearrangement that can be detected after HindIII or EcoRI digestion of genomic DNA, and is similar to that found recently in seven unrelated Hunter patients. Our data suggest that the structural aberration observed is a partial intragenic inversion. As the same altered hybridiz…
Simultaneous determination in situ of DNA fragmentation and 8-oxoguanine in human sperm.
2009
Deoxyribonucleic acid fragmentation and oxidative DNA damage were simultaneously determined in the same sperm cell, incubating with an 8-oxoguanine DNA probe on human spermatozoa processed by the sperm chromatin dispersion test. The assay was validated by incubation with agents that induce DNA fragmentation with or without oxidative base damage. In all samples examined, increased levels of 8-oxoguanine were present only in those spermatozoa with fragmented DNA, suggesting a link between both DNA damage types.
Specific DNA probes to detect Escherichia coli strains producing cytotoxic necrotising factor type 1 or type 2
1994
Cytotoxic necrotising factors type 1 (CNF1) and type 2 (CNF2) are produced by many Escherichia coli strains isolated from man and animals with intestinal or extra-intestinal colibacillosis. In most laboratories, CNF-producing strains are detected by a cell cytotoxicity assay and confirmed with a neutralisation assay or a mouse footpad assay. In this study, we sought to determine whether DNA probes could detect clinical isolates of E. coli producing CNF2 or CNF1, or both, without the need for cell cultures or animal assays. Two internal fragments of the gene encoding CNF2 were used as DNA probes: a 875-bp XhoI-PstI DNA fragment and an adjacent 335-bp PstI-ClaI fragment. A positive response w…
Enzyme-linked immunoassay for detection of PCR-amplified DNA of legionellae in bronchoalveolar fluid.
1995
A nonradioactive method is described that detects 10 to 100 legionellae in 1 ml of bronchoalveolar lavage fluid. DNA is purified by a proteinase K-phenol protocol or with a commercial DNA preparation kit and amplified by PCR with amplimers specific for the 16S rRNA gene of Legionella pneumophila. The upstream primer is 5' biotinylated. The amplification product is immobilized on streptavidin-coated microtiter plates. Because of the high binding capacity, no removal of nonincorporated biotin from the PCR product is required. After alkaline denaturation, the single-stranded PCR product is hybridized with a 5' digoxigenin-labeled probing oligomer. The amplification product is then detected by …
Characteristics of Escherichia coli strains belonging to enteropathogenic E. coli serogroups isolated in Italy from children with diarrhea.
1996
Fifty-five Escherichia coli strains belonging to enteropathogenic E. coli (EPEC) serogroups were examined for phenotypic and genetic factors associated with virulence. The strains were isolated in Italy from children with diarrhea and identified as EPEC by clinical laboratories using commercially available antisera. O:H serotyping showed that 35 strains (27 of O26, O111, and O128 serogroups) belonged to 11 serotypes considered to be classical EPEC O:H serotypes. The other 20 isolates were classified as 15 nonclassical EPEC O:H serotypes. All the potential EPEC virulence factors associated with bacterial adhesion (localized adherence, fluorescentactin staining test positivity, presence of th…
Mapping of Polytene Chromosomes
2000
Principle and Polytene chromosomes consist of up to several thousands of chromatids applications and are therefore especially suitable for direct mapping with the help ofIn situhybridization. With the introduction of nonradioactive labeling and detection methods, e.g., fluorescenceIn situhybridization (FISH) (Lan-ger-Safer et al., 1982), theIn situhybridization procedure has become easy to perform and the results can be obtained within a day. Furthermore, the method described here (Schmidt et al., 1988; Schmidt, 1992) is a simplified version which additionally allows for the hybridization of more than one DNA probe simultaneously. This double or multicolor hybridization results in very prec…
Species-specific identification of Dekkera/Brettanomyces yeasts by fluorescently labeled DNA probes targeting the 26S rRNA.
2007
Sequencing of the complete 26S rRNA genes of all Dekkera/Brettanomyces species colonizing different beverages revealed the potential for a specific primer and probe design to support diagnostic PCR approaches and FISH. By analysis of the complete 26S rRNA genes of all five currently known Dekkera/Brettanomyces species (Dekkera bruxellensis, D. anomala, Brettanomyces custersianus, B. nanus and B. naardenensis), several regions with high nucleotide sequence variability yet distinct from the D1/D2 domains were identified. FISH species-specific probes targeting the 26S rRNA gene's most variable regions were designed. Accessibility of probe targets for hybridization was facilitated by the constr…