scholarly journals Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb fromF8for simplified preimplantation genetic diagnosis of hemophilia A

2017 ◽  
Vol 15 (7) ◽  
pp. 1473-1483 ◽  
Author(s):  
M. Zhao ◽  
M. Chen ◽  
A. S. C. Tan ◽  
F. S. H. Cheah ◽  
J. Mathew ◽  
...  
Keyword(s):  
Microsatellite Markers ◽  
Preimplantation Genetic Diagnosis ◽  
Hemophilia A ◽  
Genetic Diagnosis ◽  
Single Tube ◽  
Polymorphic Microsatellite

scholarly journals Identification of Novel Microsatellite Markers <1 Mb from the HTT CAG Repeat and Development of a Single-Tube Tridecaplex PCR Panel of Highly Polymorphic Markers for Preimplantation Genetic Diagnosis of Huntington Disease

2016 ◽  
Vol 62 (8) ◽  
pp. 1096-1105 ◽  
Author(s):  
Mingjue Zhao ◽  
Min Chen ◽  
Caroline G Lee ◽  
Samuel S Chong
Keyword(s):  
Microsatellite Markers ◽  
Preimplantation Genetic Diagnosis ◽  
Huntington Disease ◽  
In Silico ◽  
Single Cells ◽  
Genetic Diagnosis ◽  
Cag Repeat ◽  
Polymorphic Markers ◽  
Single Tube ◽  
Direct Mutation

Abstract BACKGROUND Preimplantation genetic diagnosis (PGD) of Huntington disease (HD) generally employs linkage analysis of flanking microsatellite markers to complement direct mutation testing, as well as for exclusion testing. Thus far, only 10 linked markers have been developed for use in HD PGD, with a maximum of 3 markers coamplified successfully. We aimed to develop a single-tube multiplex PCR panel of highly polymorphic markers to simplify HD PGD. METHODS An in silico search was performed to identify all markers within 1 Mb flanking the huntingtin (HTT) gene. Selected markers were optimized in a single-tube PCR panel, and their polymorphism indices were determined in 2 populations. The panel was tested on 63 single cells to validate its utility in PGD. RESULTS We identified 102 markers in silico, of which 56 satisfied the selection criteria. After initial testing, 12 markers with potentially high heterozygosity were optimized into a single-tube PCR panel together with a 13th more distally located marker. Analysis of DNA from 183 Chinese and Caucasian individuals revealed high polymorphism indices for all markers (polymorphism information content &gt;0.5), with observed heterozygosities ranging from 0.5–0.92. All individuals were heterozygous for at least 5 markers, with 99.5% of individuals heterozygous for at least 2 markers upstream and downstream of the HTT CAG repeat. CONCLUSIONS The tridecaplex marker assay amplified reliably from single cells either directly or after whole genome amplification, thus validating its standalone use in HD exclusion PGD or as a complement to HTT CAG repeat expansion-mutation detection.

Single-tube nonaplex microsatellite PCR panel for preimplantation genetic diagnosis of Hb Bart's hydrops fetalis syndrome

2015 ◽  
Vol 35 (6) ◽  
pp. 534-543 ◽  
Author(s):  
Min Chen ◽  
Jerry K. Y. Chan ◽  
Sadhana Nadarajah ◽  
Arnold S. C. Tan ◽  
Melinda L. H. Chan ◽  
...  
Keyword(s):  
Preimplantation Genetic Diagnosis ◽  
Genetic Diagnosis ◽  
Hydrops Fetalis ◽  
Single Tube ◽  
Hb Bart's

scholarly journals Preimplantation genetic diagnosis for hemophilia A using indirect linkage analysis and direct genotyping approaches

2010 ◽  
Vol 8 (4) ◽  
pp. 783-789 ◽  
Author(s):  
A. D. LAURIE ◽  
A. M. HILL ◽  
J. R. HARRAWAY ◽  
A. P. FELLOWES ◽  
G. T. PHILLIPSON ◽  
...  
Keyword(s):  
Linkage Analysis ◽  
Preimplantation Genetic Diagnosis ◽  
Hemophilia A ◽  
Genetic Diagnosis ◽  
Indirect Linkage

scholarly journals Polar Body Diagnosis for Hemophilia A Using Multiplex PCR for Linked Polymorphic Markers

2005 ◽  
Vol 53 (3) ◽  
pp. 277-280 ◽  
Author(s):  
Diana Tomi ◽  
Georg Griesinger ◽  
Askan Schultze-Mosgau ◽  
Juliane Eckhold ◽  
Beate Schöpper ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Preimplantation Genetic Diagnosis ◽  
Hemophilia A ◽  
Polar Body ◽  
Genetic Diagnosis ◽  
Polymorphic Markers ◽  
Polar Bodies ◽  
Fluorescent Pcr ◽  
Polar Body Diagnosis

Preimplantation genetic diagnosis (PGD) is usually performed on blastomeres. In Germany, the only possibility to perform PGD is by analysis of polar bodies. We performed PGD using polar bodies in a woman who is a carrier of hemophilia A. Multiplex PCR followed by nested fluorescent PCR for five linked polymorphic markers was established. From 11 analyzed polar bodies, only 1 showed alleles linked to the mutation. The corresponding oocyte was transferred and no pregnancy was established. As seen in other investigations, the rate of heterozygous first polar bodies is surprisingly high.

Novel and Known Microsatellite Markers Within the β-Globin Cluster to Support Robust Preimplantation Genetic Diagnosis of β-Thalassemia and Sickle Cell Syndromes

Hemoglobin ◽  
2011 ◽  
Vol 35 (1) ◽  
pp. 56-66 ◽  
Author(s):  
Sophia Zachaki ◽  
Christina Vrettou ◽  
Aspasia Destouni ◽  
Ggeorgia Kokkali ◽  
Joanne Traeger-Synodinos ◽  
...  
Keyword(s):  
Microsatellite Markers ◽  
Sickle Cell ◽  
Preimplantation Genetic Diagnosis ◽  
Genetic Diagnosis

scholarly journals Single-Tube Dodecaplex PCR Panel of Polymorphic Microsatellite Markers Closely Linked to the DMPK CTG Repeat for Preimplantation Genetic Diagnosis of Myotonic Dystrophy Type 1

2017 ◽  
Vol 63 (6) ◽  
pp. 1127-1140 ◽  
Author(s):  
Mulias Lian ◽  
Mingjue Zhao ◽  
Caroline G Lee ◽  
Samuel S Chong
Keyword(s):  
Microsatellite Markers ◽  
Multiplex Pcr ◽  
Polymorphism Information Content ◽  
Single Cells ◽  
Genetic Diagnosis ◽  
Myotonic Dystrophy Type 1 ◽  
Myotonic Dystrophy Type ◽  
Single Tube ◽  
Ctg Repeat

Abstract BACKGROUND Preimplantation genetic diagnosis (PGD) of myotonic dystrophy type 1 (DM1) currently uses conventional PCR to detect nonexpanded dystrophia myotonica protein kinase (DMPK) alleles or triplet-primed PCR to detect the CTG-expanded alleles, coupled with analysis of linked microsatellite markers to increase diagnostic accuracy. We aimed to simplify the process of identification and selection of informative linked markers for application to DM1 PGD. METHODS An in silico search was performed to identify all markers within 1–1.5 Mb flanking the DMPK gene. Five previously known (D19S559, APOC2, D19S543, D19S112, and BV209569) and 7 novel (DM45050, DM45178, DM45209, DM45958, DM46513, DM46892, and DM47004.1) markers with potentially high heterozygosity values and polymorphism information content were selected and optimized in a single-tube multiplex PCR panel. RESULTS Analysis of 184 DNA samples of Chinese and Caucasian individuals (91 from unrelated, anonymized cord blood of Chinese babies born at the National University Hospital, Singapore, and 93 Caucasian DNA samples from the Human Variation Panel HD100CAU) confirmed the high polymorphism indices of all markers (polymorphism information content &gt;0.5), with observed heterozygosity values ranging from 0.62–0.93. All individuals were heterozygous for at least 6 markers, with 99.5% of individuals heterozygous for at least 2 markers on either side of the DMPK CTG repeat. The dodecaplex marker assay was successfully validated on 42 single cells and 12 whole genome amplified single cells. CONCLUSIONS The DM1 multiplex PCR panel is suitable for use in DM1 PGD either as a standalone linkage-based assay or as a complement to DMPK CTG repeat expansion-mutation detection.

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