DNA Diagnosis in Monogenic Diseases

1994 ◽  
Vol 10 (4) ◽  
pp. 628-643 ◽  
Author(s):  
Raye Lynn Alford ◽  
Belinda J. F. Rossiter ◽  
C. Thomas Caskey
Keyword(s):  
Single Gene ◽  
Nucleotide Sequencing ◽  
Carrier Status ◽  
Dna Diagnosis ◽  
Testing Procedures ◽  
Dna Nucleotide Sequencing ◽  
Monogenic Diseases ◽  
Allele Specific ◽  
Routine Procedures

AbstractSeveral routine procedures are available for diagnosis of diseases caused by an alteration in a single gene. These techniques include Southern analysis, the polymerase chain reaction, allele-specific oligonucleotide screening, automated DNA nucleotide sequencing, and linkage analysis. DNA testing procedures can be used for diagnosis of disease, determination of carrier status in affected families, or general screening of the population. Some of the more commonly used techniques and their applications are described in this article.

Determination of carrier status in Duchenne and Becker muscular dystrophies by quantitative polymerase chain reaction and allele-specific oligonucleotides

1990 ◽  
Vol 36 (12) ◽  
pp. 2113-2117 ◽  
Author(s):  
T W Prior ◽  
A C Papp ◽  
P J Snyder ◽  
W E Highsmith ◽  
K J Friedman ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Muscular Dystrophy ◽  
Gene Dosage ◽  
Quantitative Polymerase Chain Reaction ◽  
Becker Muscular Dystrophy ◽  
Carrier Status ◽  
Chain Reaction ◽  
Allele Specific ◽  
Polymerase Chain

Abstract Detection of carriers of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD), in the deletion cases, involves calculating gene dosage from Southern blots. We show that the analysis of dosage can also be made from the polymerase chain reaction (PCR) with use of allele-specific oligonucleotides (ASOs). The deletion-prone exons are amplified, transferred to a membrane, and hybridized with ASOs complementary to the exons; the autoradiographic bands are then quantified with a densitometer. After determining the quantitative conditions of the amplification reaction, we were able to identify deletions in a DMD/BMD carrier female. The determination of carrier status via PCR removes several of the technical limitations of Southern analysis and is also cost- and labor-effective.

Determination of haemophilia A carrier status by mutation analysis

Haemophilia ◽  
2001 ◽  
Vol 7 (1) ◽  
pp. 20-25 ◽  
Author(s):  
S. Oranwiroon ◽  
V. Akkarapatumwong ◽  
P. Pung-Amritt ◽  
A. Treesucon ◽  
G. Veerakul ◽  
...  
Keyword(s):  
Mutation Analysis ◽  
Carrier Status ◽  
Haemophilia A

scholarly journals Studies on Colchicine Induced Chromosome Doubling for Enhancement of Quality Traits in Ornamental Plants

Plants ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 194 ◽  
Author(s):  
Ayesha Manzoor ◽  
Touqeer Ahmad ◽  
Muhammad Bashir ◽  
Ishfaq Hafiz ◽  
Cristian Silvestri
Keyword(s):  
Chromosome Doubling ◽  
Ornamental Plants ◽  
Plant Size ◽  
Unreduced Gametes ◽  
Chromosome Counting ◽  
Morphological And Physiological Traits ◽  
Routine Procedures ◽  
Mitotic Inhibitor ◽  
Intense Color

Polyploidy has the utmost importance in horticulture for the development of new ornamental varieties with desirable morphological traits referring to plant size and vigor, leaf thickness, larger flowers with thicker petals, intense color of leaves and flowers, long lasting flowers, compactness, dwarfness and restored fertility. Polyploidy may occur naturally due to the formation of unreduced gametes or can be artificially induced by doubling the number of chromosomes in somatic cells. Usually, natural polyploid plants are unavailable, so polyploidy is induced synthetically with the help of mitotic inhibitors. Colchicine is a widely used mitotic inhibitor for the induction of polyploidy in plants during their cell division by inhibiting the chromosome segregation. Different plant organs like seeds, apical meristems, flower buds, and roots can be used to induce polyploidy through many application methods such as dipping/soaking, dropping or cotton wool. Flow cytometry and chromosome counting, with an observation of morphological and physiological traits are routine procedures for the determination of ploidy level in plants.

scholarly journals Copy number variation in MODY diabetes - Familial case presentation

2018 ◽  
Vol 2 (2) ◽  
pp. 73
Author(s):  
Naida Lojo-Kadric ◽  
Zelija Velija Asimi ◽  
Jasmin Ramic ◽  
Ksenija Radic ◽  
Lejla Pojskic
Keyword(s):  
Insulin Secretion ◽  
Copy Number ◽  
Autosomal Dominant ◽  
Single Gene ◽  
Maturity Onset Diabetes ◽  
Dominant Form ◽  
Case Presentation ◽  
Monogenic Diseases ◽  
Number Variation ◽  
Autosomal Dominant Form

MODY (maturity-onset diabetes of the young) is an autosomal dominant form of diabetes that is usually manifested before the 25-year of life. This type of diabetes is caused by defects in the primary insulin secretion. There are several types of MODY, which are monogenic diseases, where mutations in a single gene are responsible for a particular type of MODY. Currently, there are eleven types of MODY, from which the most common types are MODY 2 and MODY 3 (with mutations on GCK and HNF1A genes, respectively). We identified very rare MODY 7 type of diabetes in three family members by MLPA analysis.

scholarly journals Four Ia invariant chain forms derive from a single gene by alternate splicing and alternate initiation of transcription/translation.

1987 ◽  
Vol 166 (2) ◽  
pp. 444-460 ◽  
Author(s):  
D M O'Sullivan ◽  
D Noonan ◽  
V Quaranta
Keyword(s):  
Single Gene ◽  
Structural Basis ◽  
Invariant Chain ◽  
Nucleotide Sequencing ◽  
S1 Nuclease ◽  
Initiation Of Translation ◽  
Associated Proteins ◽  
Alternatively Spliced ◽  
Translational Start ◽  
Genomic Regions

We determined the structural basis for the presence of electrophoretically-distinct, antigenically-related forms of invariant chains in Ia oligomers, and established the mechanisms by which they can be expressed from a single gene. S1 nuclease protection assays indicated that, in B cells, transcription of this gene initiates at a minimum of three sites. Thus, unlike previously thought, invariant chain mRNAs have heterogeneous 5' untranslated segments that may differentially affect initiation of translation. Further, restriction mapping and nucleotide sequencing of cDNAs revealed two kinds of invariant chain mRNAs differing by an internal coding segment of 192 bp. This segment represents an alternatively spliced exon, as demonstrated by nucleotide sequencing of corresponding genomic regions. The exon (exon X) encodes a cysteine-rich stretch of 64 amino acids near the COOH terminus that displays a striking and surprising homology to an internal amino acid repeat of thyroglobulin, suggesting an evolutionary mechanism of exon shuffling. Transient expression of cDNAs indicated that both types of alternatively spliced mRNAs contain two in-frame AUGs functioning as alternate start sites for translation. Thus, transfections with exon X-lacking cDNAs resulted in the expression of Mr 33,000 and 31,000 proteins, detected by immunoprecipitation with anti-invariant chain antisera, and identical by two-dimensional gel (2-D) analyses to the B cell invariant-chain forms gamma 1 (Mr 31,000), gamma 2, and gamma 3 (Mr 33,000). Similarly, exon X-containing cDNAs expressed Mr 43,000 and 41,000 proteins, also identical by 2-D migration to Ia-associated proteins. Thus, human Ia molecules contain four forms of invariant chain of closely related but nonidentical primary structure that are generated from a single gene by a complex pattern of alternate transcriptional start, exon splicing, and translational start.

scholarly journals Use of a BamHI polymorphism in the factor IX gene for the determination of hemophilia B carrier status

Blood ◽  
1986 ◽  
Vol 67 (5) ◽  
pp. 1508-1511 ◽  
Author(s):  
CW Hay ◽  
KA Robertson ◽  
SL Yong ◽  
AR Thompson ◽  
GH Growe ◽  
...  
Keyword(s):  
Human Factor ◽  
Factor Ix ◽  
Hemophilia B ◽  
Carrier Status ◽  
X Chromosomes ◽  
Human Factor Ix ◽  
History Of

Abstract A BamHI polymorphism has been identified in the human factor IX gene. This polymorphism, which occurs in approximately 6% of X chromosomes, has been used to determine the carrier status of a female in a family with a history of hemophilia B. This family was uninformative for the previously reported TaqI and Xmnl polymorphisms in the factor IX gene.

scholarly journals Determination of RhD Zygosity: Comparison of a Double Amplification Refractory Mutation System Approach and a Multiplex Real-Time Quantitative PCR Approach

2001 ◽  
Vol 47 (4) ◽  
pp. 667-672 ◽  
Author(s):  
Rossa W K Chiu ◽  
Michael F Murphy ◽  
Carrie Fidler ◽  
Benny C Y Zee ◽  
James S Wainscoat ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Gene Dosage ◽  
Amplification Refractory Mutation System ◽  
Pcr Assay ◽  
Real Time Quantitative Pcr ◽  
Mutation System ◽  
Allele Specific ◽  
Quantitative Pcr Assay

Abstract Background: Rh isoimmunization and hemolytic disease of the newborn still occur despite the availability of Rh immunoglobulin. For the prenatal investigation of sensitized RhD-negative pregnant women, determination of the zygosity of the RhD-positive father has important implications. The currently available molecular methods for RhD zygosity assessment, in general, are technically demanding and labor-intensive. Therefore, at present, rhesus genotype assessment is most commonly inferred from results of serological tests. The recent elucidation of the genetic structure of the prevalent RHD deletion in Caucasians, as well as the development of real-time PCR, allowed us to explore two new approaches for the molecular determination of RhD zygosity. Methods: Two methods for RhD zygosity determination were developed. The first was based on the double Amplification Refractory Mutation System (double ARMS). The second was based on multiplex real-time quantitative PCR. For the double ARMS assay, allele-specific primers were designed to directly amplify the most prevalent RHD deletion found in RhD-negative individuals in the Caucasian population. The multiplex real-time quantitative PCR assay, on the other hand, involved coamplification and quantification of RHD-specific sequences in relation to a reference gene, albumin, in a single PCR reaction. A ratio, ΔCt, based on the threshold cycle, was then determined and reflects the RHD gene dosage. Results: The allele-specific primers of the double ARMS assay reliably amplified the RHD-deleted allele and therefore accurately distinguished homozygous from heterozygous RhD-positive samples. The results were in complete concordance with serological testing. For the multiplex real-time quantitative PCR assay, the ΔCt values clearly segregated into two distinct populations according to the RHD gene dosage, with mean values of 1.70 (SD, 0.17) and 2.62 (SD, 0.29) for the homozygous and heterozygous samples, respectively (P <0.001, t-test). The results were in complete concordance with the results of serological testing as well as with the double ARMS assay. Conclusion: Double ARMS and real-time quantitative PCR are alternative robust assays for the determination of RhD zygosity.

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