Frequency of the cystic fibrosis mutation delta F508 in Poland.
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Cystic Fibrosis↗
Biomedical subjects
J Reiss
Publications and source records attributed to J Reiss.
Detection of biotoxic compounds with the 2,3,5-triphenyltetrazolium chloride seed germination test.
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Copper↗
Prenatal deletion detection in a sporadic case of Duchenne muscular dystrophy without genotype information from the affected individual.
We describe the application of deletion screening by amplification of deletion-prone exons via polymerase chain reaction (PCR) in a family with a sporadic case of Duchenne muscular dystrophy (DMD). No DNA was available from the affected patient who died 12 years beforehand at the age of 18 years. Material obtained prenatally from two male fetuses exhibited an identical deletion. These findings effectively transformed a sporadic case into a familial case and a numerical carrier risk was substituted by obligate carrier status. Additionally an indirect genotype analysis was replaced by the possibility of direct DNA analysis. Genetic counselling, formerly based upon incomplete data, can now be aided by precise risk assessment.
Adult↗
The polymerase chain reaction and its potential role in clinical diagnostics and research.
In-vitro amplification of deoxyribonucleic acid molecules by means of the polymerase chain reaction (PCR) must be regarded as the most important advance in the life sciences to take place during the last decade. Originally applied to the identification of mutations in well-known and fully sequenced human genes, its applications have now been extended to a wide variety of biological and medical disciplines, accompanied by significant technical improvements and sophisticated variations of the basic principle. Specialized molecular genetics laboratories were the first to employ this new method, and they still are in the process of extending its potential. Due to its unique properties, applications of PCR quickly spread to other areas of research, and numerous clinical studies have already employed PCR. The field is currently still expanding rapidly.
Forecasting↗
A cystic fibrosis patient homozygous for the nonsense mutation R553X.
A cystic fibrosis patient homozygous for the nonsense mutation R553X was identified by mutation screening and the genotype confirmed by DNA sequencing. This patient, the only one described to date who is homozygous for this stop codon in exon 11 of the CFTR gene, is moderately severely affected. Clinical and molecular findings are presented.
Alleles↗
A cystic fibrosis patient with the nonsense mutation G542X and the splice site mutation 1717-1.
A cystic fibrosis patient with the genotype G542X/1717-1 (G----A) was identified by DNA sequencing of exon 11 of the CFTR gene. The available molecular and clinical data are presented. This is the first report of a patient with this rare genotype and may serve to improve our understanding of allele interactions.
Amino Acid Sequence↗
[A simplified method of rapid hemagglutination test for determining the levels of tetanus antibodies].
A new, simplified methodology of preparation of lyophilized chrome-coated turkey blood cells (formalin-treated ) for rapid identification of tetanus antibodies in indirect hemagglutination test is described. Blood cells diagnostic preparations obtained in this way were easier and three times faster to prepare than tanned cell preparations . They maintained unchanged capability for specific agglutination during at least one year of storage when kept at 4 degrees C. proposed methodology enable to start a production of laboratory kits necessary for controlled prophylaxis of tetanus.
Animals↗
[Principles of DNA diagnosis exemplified by cystic fibrosis].
Principles of genetic disease analysis using molecular genetic techniques are illustrated. Direct and indirect genotyping is described with cystic fibrosis as an example. The possibilities to combine genetic markers for "full informativity" are included.
Child↗
The effect of replication errors on the mismatch analysis of PCR-amplified DNA.
The mismatch analysis of PCR-amplified DNA has generally assumed the absence of artificially introduced base substitutions in a significant proportion of the amplification product. This technique, however, differs from the direct sequencing of amplified DNA in that non-specific substitutions will render a molecule useless in analysis. The expected signal-to-noise ratio is heavily influenced by several parameters viz. initial template copy number, number of replication cycles, eventual product yield and the type of experimental system adopted. Mathematical modelling can be used to optimize fragment length with respect to the method applied and suggests as yet undescribed improvements such as partial modification or cleavage to optimize signal detection.
Base Composition↗
Detection of a novel point mutation causing haemophilia A by PCR/direct sequencing of ectopically-transcribed factor VIII mRNA.
Specifically-primed reverse transcripts of lymphocyte-derived factor VIII (FVIII) mRNA were successfully amplified by means of the polymerase chain reaction (PCR) thus further extending the phenomenon of ectopic ("illegitimate") transcription of tissue-specific genes. The diagnostic potential of a basal rate of transcription in non-expressing tissues was then demonstrated by the detection of a novel point mutation in the FVIII gene causing haemophilia A by PCR/direct sequencing of ectopically transcribed mRNA derived from patient lymphocytes.
Base Sequence↗
Rapid detection of deletions in the Duchenne muscular dystrophy gene by PCR amplification of deletion-prone exon sequences.
Using the polymerase chain reaction (PCR) technique, we have screened the DNA of 42 patients with Duchenne or Becker muscular dystrophy for deletions within the DMD gene. Two regions within putative deletion "hot spots" of this gene were tested, and deletions were found in 16.6% of patients. The oligonucleotide primers employed in this study initiate the amplification of exon sequences and were used to test the suitability and reliability of PCR in deletion screening and prenatal diagnosis using various numbers of cycles and artificial contamination ratios. We compared our approach with both "multiplex DNA amplification" and Southern blot analysis. A comparative evaluation of currently available techniques is presented.
Base Sequence↗
Application of the polymerase chain reaction to the diagnosis of human genetic disease.
In vitro DNA amplification by means of the polymerase chain reaction is currently revolutionizing human molecular genetics. Since its inception in 1985, a wide variety of different methods and their applications in the diagnosis of disease have been described. This review is intended to serve as a brief guide to current and emerging possibilities in this rapidly expanding field.
Anemia, Sickle Cell↗
Diagnosis of haemophilia B using the polymerase chain reaction.
The polymerase chain reaction (PCR) was used to amplify specific DNA sequences within the factor IX gene of haemophilia B patients and their relatives. Three of the amplified fragments contain polymorphic sites, which can be used as markers in segregation analyses. These restriction fragment length polymorphisms (RFLPs) were until recently detected by Southern blotting after digestion with the restriction enzymes Taq I, Dde I and Xmn I. All three RFLP's are located in introns of the factor IX gene and together are informative in approximately 70% of all cases. Each of the polymorphisms was successfully used in carrier detection studies after amplification of the relevant fragments. This method is also suitable for rapid antenatal diagnosis. Additionally we were able to amplify all eight exons of the factor IX gene including the splice junctions and a part of the 5'-region. Large deletions or insertions can be detected without further analysis. Several possibilities for the rapid detection of point mutations after DNA amplification have been described recently. The complete amplification of all functional parts of the Factor IX gene in combination with these new techniques should enable us to detect the majority of mutations leading to haemophilia B.
Base Sequence↗
Frequency of the F508 deletion in the CFTR gene in Turkish cystic fibrosis patients.
The F508 deletion in the cystic fibrosis transmembrane conductance regulator (CFTR) gene was found in 8 out of 30 Turkish cystic fibrosis (CF) chromosomes (27%). Five Turkish delta F508 CF chromosomes were associated with the risk haplotype B in KM19 (2 allele)/XV2c (1 allele). In the Turkish population, cystic fibrosis is predominantly caused by mutations other than the F508 deletion.
Chromosome Deletion↗
Characterization of pathological dystrophin transcripts from the lymphocytes of a muscular dystrophy carrier.
Both normal and pathological transcripts of tissue-specific genes may be detected by polymerase chain reaction (PCR) amplification in tissues not normally considered to express the gene product. The exploitation of constitutive basal mRNA levels ("ectopic" transcription) would be a major boon to diagnostic medicine since it promises both to simplify the analysis of complex genes and to avoid the requirement for an expressing tissue that is sometimes obtainable only by biopsy. We have demonstrated the feasibility of this novel strategy by characterizing a mutation in the X-chromosomal Duchenne (or Becker) muscular dystrophy (DMD/BMD) gene encoding dystrophin. The massive size of this gene has in the past often hindered carrier detection due to the high frequency of recombination and the high proportion of new mutations. In this study a deletion was identified in both a BMD patient and a heterozygous carrier using only a minimal volume of peripheral blood. Following specifically primed reverse transcription of lymphocyte RNA, the relevant region of the pathological cDNA was PCR-amplified. Sequence analysis indicated an in-frame deletion of exons 45 to 47.
DNA↗
Direct gene diagnosis of cystic fibrosis by allele-specific polymerase chain reactions.
Cloning of the cystic fibrosis gene and the identification of the predominant disease-causing mutation did not only help in the understanding of this frequent disease, but was immediately followed by applications in direct gene diagnosis. We describe a method for the detection of the so-called delta F508 deletion, which accounts for 70% of the mutations: a polymerase chain reaction with two different combinations of oligonucleotide primers, which discriminate between mutant and wild-type alleles. This allele-specific amplification provides a rapid, non-radioactive and very reliable method for direct genotyping. Establishment of the procedure and its application in diagnosis are described. We further report preliminary data on the frequency of this mutation in German patients and its association with restriction fragment length polymorphism haplotypes.
Alleles↗
[Direct genetic diagnosis in cystic fibrosis].
The recent cloning of the cystic fibrosis gene and the simultaneous identification of the predominant mutation enabled direct gene diagnosis in the majority of all CF families. We demonstrate the superiority of now available direct genotyping in five individual cases.
Adult↗
[Papaverin--effective in therapy of pruritus of atopic dermatitis?].
Patients suffering from atopic dermatitis show increased phosphodiesterase activity in leucocytes. As a result, the level of intracellular c-AMP is reduced, which can produce a number of metabolic dysfunctions: We found strong evidence that an enhanced release of several inflammation mediators can favor itching, which in turn promotes the development of eczema, since it induces reflectory scratching. In order to solve the question whether papaverine as a phosphodiesterase inhibitor might offer a new approach in the systemic therapy of itching, we performed a placebo-controlled study on 51 patients with atopic dermatitis, who were orally treated with papaverin-HCl over 8 weeks. However, we did not observe any improvement of itching or eczema during treatment. Some other recently developed phosphodiesterase inhibitors are discussed.
Administration, Oral↗