Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A
Essentials Preimplantation genetic diagnosis (PGD) of severe hemophilia A relies on linkage analysis. Simultaneous multi‐marker screening can simplify selection of informative markers in a couple. We developed a single‐tube tetradecaplex panel of polymorphic markers for hemophilia A PGD use. Informa...
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| Veröffentlicht in: | Journal of thrombosis and haemostasis 2017-07, Vol.15 (7), p.1473-1483 |
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| creator | Zhao, M. Chen, M. Tan, A. S. C. Cheah, F. S. H. Mathew, J. Wong, P. C. Chong, S. S. |
| description | Essentials
Preimplantation genetic diagnosis (PGD) of severe hemophilia A relies on linkage analysis.
Simultaneous multi‐marker screening can simplify selection of informative markers in a couple.
We developed a single‐tube tetradecaplex panel of polymorphic markers for hemophilia A PGD use.
Informative markers can be used for linkage analysis alone or combined with mutation detection.
Summary
Background
It is currently not possible to perform single‐cell preimplantation genetic diagnosis (PGD) to directly detect the common inversion mutations of the factor VIII (F8) gene responsible for severe hemophilia A (HEMA). As such, PGD for such inversion carriers relies on indirect analysis of linked polymorphic markers.
Objectives
To simplify linkage‐based PGD of HEMA, we aimed to develop a panel of highly polymorphic microsatellite markers located near the F8 gene that could be simultaneously genotyped in a multiplex‐PCR reaction.
Methods
We assessed the polymorphism of various microsatellite markers located ≤ 1 Mb from F8 in 177 female subjects. Highly polymorphic markers were selected for co‐amplification with the AMELX/Y indel dimorphism in a single‐tube reaction.
Results
Thirteen microsatellite markers located within 0.6 Mb of F8 were successfully co‐amplified with AMELX/Y in a single‐tube reaction. Observed heterozygosities of component markers ranged from 0.43 to 0.84, and ∼70–80% of individuals were heterozygous for ≥ 5 markers. The tetradecaplex panel successfully identified fully informative markers in a couple interested in PGD for HEMA because of an intragenic F8 point mutation, with haplotype phasing established through a carrier daughter. In‐vitro fertilization (IVF)‐PGD involved single‐tube co‐amplification of fully informative markers with AMELX/Y and the mutation‐containing F8 amplicon, followed by microsatellite analysis and amplicon mutation‐site minisequencing analysis.
Conclusions
The single‐tube multiplex‐PCR format of this highly polymorphic microsatellite marker panel simplifies identification and selection of informative markers for linkage‐based PGD of HEMA. Informative markers can also be easily co‐amplified with mutation‐containing F8 amplicons for combined mutation detection and linkage analysis. |
| doi_str_mv | 10.1111/jth.13685 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_journals_1915256512</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1915256512</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3885-149ef3487ad126370472fcda36c8366c1ade4d12e03f9325c80dd2c1b69ebcb43</originalsourceid><addsrcrecordid>eNp1kcFO3DAURa2KqlDogh9AT2LFYiC244wjsUEISiuqLqDryHGeZzw4cbA9gtnxCf2CfhxfgocBdvXGfvLRvbrvErJPi2Oaz8kizY8pr6T4RHao4HIylbzaen_XnG-TrzEuioLWghVfyDaTvBRMyh3y78YOM4fPT3_TskVImILqUKvR4SOMakAH3sDczuZuBaN3q96HcW419FYHH1VC52xC6FW4wxDhFCj8asEE38OlBOMDRNuPzhqLHYwB14MakkrWDzDDAVMW66yaDT7a-GqGvc8Wzio42yOfjXIRv73du-TP5cXt-dXk-vf3H-dn1xPNpRQTWtZoeCmnqqOs4tOinDKjO8UrnTdRaZozlfkLC25qzoSWRdcxTduqxla3Jd8lhxvdMfj7JcbULPwyDNmyoTUVTFSCskwdbah19BjQNGOwOfmqoUWzbqLJTTSvTWT24E1x2fbYfZDvq8_AyQZ4sA5X_1dqft5ebSRfAN31ll0</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1915256512</pqid></control><display><type>article</type><title>Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>Zhao, M. ; Chen, M. ; Tan, A. S. C. ; Cheah, F. S. H. ; Mathew, J. ; Wong, P. C. ; Chong, S. S.</creator><creatorcontrib>Zhao, M. ; Chen, M. ; Tan, A. S. C. ; Cheah, F. S. H. ; Mathew, J. ; Wong, P. C. ; Chong, S. S.</creatorcontrib><description>Essentials
Preimplantation genetic diagnosis (PGD) of severe hemophilia A relies on linkage analysis.
Simultaneous multi‐marker screening can simplify selection of informative markers in a couple.
We developed a single‐tube tetradecaplex panel of polymorphic markers for hemophilia A PGD use.
Informative markers can be used for linkage analysis alone or combined with mutation detection.
Summary
Background
It is currently not possible to perform single‐cell preimplantation genetic diagnosis (PGD) to directly detect the common inversion mutations of the factor VIII (F8) gene responsible for severe hemophilia A (HEMA). As such, PGD for such inversion carriers relies on indirect analysis of linked polymorphic markers.
Objectives
To simplify linkage‐based PGD of HEMA, we aimed to develop a panel of highly polymorphic microsatellite markers located near the F8 gene that could be simultaneously genotyped in a multiplex‐PCR reaction.
Methods
We assessed the polymorphism of various microsatellite markers located ≤ 1 Mb from F8 in 177 female subjects. Highly polymorphic markers were selected for co‐amplification with the AMELX/Y indel dimorphism in a single‐tube reaction.
Results
Thirteen microsatellite markers located within 0.6 Mb of F8 were successfully co‐amplified with AMELX/Y in a single‐tube reaction. Observed heterozygosities of component markers ranged from 0.43 to 0.84, and ∼70–80% of individuals were heterozygous for ≥ 5 markers. The tetradecaplex panel successfully identified fully informative markers in a couple interested in PGD for HEMA because of an intragenic F8 point mutation, with haplotype phasing established through a carrier daughter. In‐vitro fertilization (IVF)‐PGD involved single‐tube co‐amplification of fully informative markers with AMELX/Y and the mutation‐containing F8 amplicon, followed by microsatellite analysis and amplicon mutation‐site minisequencing analysis.
Conclusions
The single‐tube multiplex‐PCR format of this highly polymorphic microsatellite marker panel simplifies identification and selection of informative markers for linkage‐based PGD of HEMA. Informative markers can also be easily co‐amplified with mutation‐containing F8 amplicons for combined mutation detection and linkage analysis.</description><identifier>ISSN: 1538-7933</identifier><identifier>ISSN: 1538-7836</identifier><identifier>EISSN: 1538-7836</identifier><identifier>DOI: 10.1111/jth.13685</identifier><identifier>PMID: 28345288</identifier><language>eng</language><publisher>England: Elsevier Limited</publisher><subject>Alleles ; Coagulation factors ; Diagnosis ; Dimorphism ; DNA Mutational Analysis ; Electrophoresis, Capillary ; Exons ; Factor VIII - genetics ; Factor VIII deficiency ; Female ; Fertilization in Vitro ; Gene loci ; Genetic Linkage ; Genetic Markers ; Genetic screening ; Genotype ; Haplotypes ; Hemophilia ; hemophilia A ; Hemophilia A - blood ; Hemophilia A - diagnosis ; Hemophilia A - genetics ; Heterozygote ; Humans ; In vitro fertilization ; INDEL Mutation ; Inversion ; Linkage analysis ; Male ; microsatellite ; Microsatellite Repeats ; Microsatellites ; Mutation ; Oocytes - metabolism ; Point Mutation ; Polymerase chain reaction ; Polymorphism, Genetic ; Pregnancy ; Preimplantation Diagnosis ; preimplantation genetic diagnosis ; Sequence Analysis, DNA ; short tandem repeat</subject><ispartof>Journal of thrombosis and haemostasis, 2017-07, Vol.15 (7), p.1473-1483</ispartof><rights>2017 International Society on Thrombosis and Haemostasis</rights><rights>2017 International Society on Thrombosis and Haemostasis.</rights><rights>Copyright © 2017 International Society on Thrombosis and Haemostasis</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3885-149ef3487ad126370472fcda36c8366c1ade4d12e03f9325c80dd2c1b69ebcb43</citedby><cites>FETCH-LOGICAL-c3885-149ef3487ad126370472fcda36c8366c1ade4d12e03f9325c80dd2c1b69ebcb43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28345288$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhao, M.</creatorcontrib><creatorcontrib>Chen, M.</creatorcontrib><creatorcontrib>Tan, A. S. C.</creatorcontrib><creatorcontrib>Cheah, F. S. H.</creatorcontrib><creatorcontrib>Mathew, J.</creatorcontrib><creatorcontrib>Wong, P. C.</creatorcontrib><creatorcontrib>Chong, S. S.</creatorcontrib><title>Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A</title><title>Journal of thrombosis and haemostasis</title><addtitle>J Thromb Haemost</addtitle><description>Essentials
Preimplantation genetic diagnosis (PGD) of severe hemophilia A relies on linkage analysis.
Simultaneous multi‐marker screening can simplify selection of informative markers in a couple.
We developed a single‐tube tetradecaplex panel of polymorphic markers for hemophilia A PGD use.
Informative markers can be used for linkage analysis alone or combined with mutation detection.
Summary
Background
It is currently not possible to perform single‐cell preimplantation genetic diagnosis (PGD) to directly detect the common inversion mutations of the factor VIII (F8) gene responsible for severe hemophilia A (HEMA). As such, PGD for such inversion carriers relies on indirect analysis of linked polymorphic markers.
Objectives
To simplify linkage‐based PGD of HEMA, we aimed to develop a panel of highly polymorphic microsatellite markers located near the F8 gene that could be simultaneously genotyped in a multiplex‐PCR reaction.
Methods
We assessed the polymorphism of various microsatellite markers located ≤ 1 Mb from F8 in 177 female subjects. Highly polymorphic markers were selected for co‐amplification with the AMELX/Y indel dimorphism in a single‐tube reaction.
Results
Thirteen microsatellite markers located within 0.6 Mb of F8 were successfully co‐amplified with AMELX/Y in a single‐tube reaction. Observed heterozygosities of component markers ranged from 0.43 to 0.84, and ∼70–80% of individuals were heterozygous for ≥ 5 markers. The tetradecaplex panel successfully identified fully informative markers in a couple interested in PGD for HEMA because of an intragenic F8 point mutation, with haplotype phasing established through a carrier daughter. In‐vitro fertilization (IVF)‐PGD involved single‐tube co‐amplification of fully informative markers with AMELX/Y and the mutation‐containing F8 amplicon, followed by microsatellite analysis and amplicon mutation‐site minisequencing analysis.
Conclusions
The single‐tube multiplex‐PCR format of this highly polymorphic microsatellite marker panel simplifies identification and selection of informative markers for linkage‐based PGD of HEMA. Informative markers can also be easily co‐amplified with mutation‐containing F8 amplicons for combined mutation detection and linkage analysis.</description><subject>Alleles</subject><subject>Coagulation factors</subject><subject>Diagnosis</subject><subject>Dimorphism</subject><subject>DNA Mutational Analysis</subject><subject>Electrophoresis, Capillary</subject><subject>Exons</subject><subject>Factor VIII - genetics</subject><subject>Factor VIII deficiency</subject><subject>Female</subject><subject>Fertilization in Vitro</subject><subject>Gene loci</subject><subject>Genetic Linkage</subject><subject>Genetic Markers</subject><subject>Genetic screening</subject><subject>Genotype</subject><subject>Haplotypes</subject><subject>Hemophilia</subject><subject>hemophilia A</subject><subject>Hemophilia A - blood</subject><subject>Hemophilia A - diagnosis</subject><subject>Hemophilia A - genetics</subject><subject>Heterozygote</subject><subject>Humans</subject><subject>In vitro fertilization</subject><subject>INDEL Mutation</subject><subject>Inversion</subject><subject>Linkage analysis</subject><subject>Male</subject><subject>microsatellite</subject><subject>Microsatellite Repeats</subject><subject>Microsatellites</subject><subject>Mutation</subject><subject>Oocytes - metabolism</subject><subject>Point Mutation</subject><subject>Polymerase chain reaction</subject><subject>Polymorphism, Genetic</subject><subject>Pregnancy</subject><subject>Preimplantation Diagnosis</subject><subject>preimplantation genetic diagnosis</subject><subject>Sequence Analysis, DNA</subject><subject>short tandem repeat</subject><issn>1538-7933</issn><issn>1538-7836</issn><issn>1538-7836</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kcFO3DAURa2KqlDogh9AT2LFYiC244wjsUEISiuqLqDryHGeZzw4cbA9gtnxCf2CfhxfgocBdvXGfvLRvbrvErJPi2Oaz8kizY8pr6T4RHao4HIylbzaen_XnG-TrzEuioLWghVfyDaTvBRMyh3y78YOM4fPT3_TskVImILqUKvR4SOMakAH3sDczuZuBaN3q96HcW419FYHH1VC52xC6FW4wxDhFCj8asEE38OlBOMDRNuPzhqLHYwB14MakkrWDzDDAVMW66yaDT7a-GqGvc8Wzio42yOfjXIRv73du-TP5cXt-dXk-vf3H-dn1xPNpRQTWtZoeCmnqqOs4tOinDKjO8UrnTdRaZozlfkLC25qzoSWRdcxTduqxla3Jd8lhxvdMfj7JcbULPwyDNmyoTUVTFSCskwdbah19BjQNGOwOfmqoUWzbqLJTTSvTWT24E1x2fbYfZDvq8_AyQZ4sA5X_1dqft5ebSRfAN31ll0</recordid><startdate>201707</startdate><enddate>201707</enddate><creator>Zhao, M.</creator><creator>Chen, M.</creator><creator>Tan, A. S. C.</creator><creator>Cheah, F. S. H.</creator><creator>Mathew, J.</creator><creator>Wong, P. C.</creator><creator>Chong, S. S.</creator><general>Elsevier Limited</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7T5</scope><scope>H94</scope><scope>K9.</scope></search><sort><creationdate>201707</creationdate><title>Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A</title><author>Zhao, M. ; Chen, M. ; Tan, A. S. C. ; Cheah, F. S. H. ; Mathew, J. ; Wong, P. C. ; Chong, S. S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3885-149ef3487ad126370472fcda36c8366c1ade4d12e03f9325c80dd2c1b69ebcb43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Alleles</topic><topic>Coagulation factors</topic><topic>Diagnosis</topic><topic>Dimorphism</topic><topic>DNA Mutational Analysis</topic><topic>Electrophoresis, Capillary</topic><topic>Exons</topic><topic>Factor VIII - genetics</topic><topic>Factor VIII deficiency</topic><topic>Female</topic><topic>Fertilization in Vitro</topic><topic>Gene loci</topic><topic>Genetic Linkage</topic><topic>Genetic Markers</topic><topic>Genetic screening</topic><topic>Genotype</topic><topic>Haplotypes</topic><topic>Hemophilia</topic><topic>hemophilia A</topic><topic>Hemophilia A - blood</topic><topic>Hemophilia A - diagnosis</topic><topic>Hemophilia A - genetics</topic><topic>Heterozygote</topic><topic>Humans</topic><topic>In vitro fertilization</topic><topic>INDEL Mutation</topic><topic>Inversion</topic><topic>Linkage analysis</topic><topic>Male</topic><topic>microsatellite</topic><topic>Microsatellite Repeats</topic><topic>Microsatellites</topic><topic>Mutation</topic><topic>Oocytes - metabolism</topic><topic>Point Mutation</topic><topic>Polymerase chain reaction</topic><topic>Polymorphism, Genetic</topic><topic>Pregnancy</topic><topic>Preimplantation Diagnosis</topic><topic>preimplantation genetic diagnosis</topic><topic>Sequence Analysis, DNA</topic><topic>short tandem repeat</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhao, M.</creatorcontrib><creatorcontrib>Chen, M.</creatorcontrib><creatorcontrib>Tan, A. S. C.</creatorcontrib><creatorcontrib>Cheah, F. S. H.</creatorcontrib><creatorcontrib>Mathew, J.</creatorcontrib><creatorcontrib>Wong, P. C.</creatorcontrib><creatorcontrib>Chong, S. S.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><jtitle>Journal of thrombosis and haemostasis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhao, M.</au><au>Chen, M.</au><au>Tan, A. S. C.</au><au>Cheah, F. S. H.</au><au>Mathew, J.</au><au>Wong, P. C.</au><au>Chong, S. S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A</atitle><jtitle>Journal of thrombosis and haemostasis</jtitle><addtitle>J Thromb Haemost</addtitle><date>2017-07</date><risdate>2017</risdate><volume>15</volume><issue>7</issue><spage>1473</spage><epage>1483</epage><pages>1473-1483</pages><issn>1538-7933</issn><issn>1538-7836</issn><eissn>1538-7836</eissn><abstract>Essentials
Preimplantation genetic diagnosis (PGD) of severe hemophilia A relies on linkage analysis.
Simultaneous multi‐marker screening can simplify selection of informative markers in a couple.
We developed a single‐tube tetradecaplex panel of polymorphic markers for hemophilia A PGD use.
Informative markers can be used for linkage analysis alone or combined with mutation detection.
Summary
Background
It is currently not possible to perform single‐cell preimplantation genetic diagnosis (PGD) to directly detect the common inversion mutations of the factor VIII (F8) gene responsible for severe hemophilia A (HEMA). As such, PGD for such inversion carriers relies on indirect analysis of linked polymorphic markers.
Objectives
To simplify linkage‐based PGD of HEMA, we aimed to develop a panel of highly polymorphic microsatellite markers located near the F8 gene that could be simultaneously genotyped in a multiplex‐PCR reaction.
Methods
We assessed the polymorphism of various microsatellite markers located ≤ 1 Mb from F8 in 177 female subjects. Highly polymorphic markers were selected for co‐amplification with the AMELX/Y indel dimorphism in a single‐tube reaction.
Results
Thirteen microsatellite markers located within 0.6 Mb of F8 were successfully co‐amplified with AMELX/Y in a single‐tube reaction. Observed heterozygosities of component markers ranged from 0.43 to 0.84, and ∼70–80% of individuals were heterozygous for ≥ 5 markers. The tetradecaplex panel successfully identified fully informative markers in a couple interested in PGD for HEMA because of an intragenic F8 point mutation, with haplotype phasing established through a carrier daughter. In‐vitro fertilization (IVF)‐PGD involved single‐tube co‐amplification of fully informative markers with AMELX/Y and the mutation‐containing F8 amplicon, followed by microsatellite analysis and amplicon mutation‐site minisequencing analysis.
Conclusions
The single‐tube multiplex‐PCR format of this highly polymorphic microsatellite marker panel simplifies identification and selection of informative markers for linkage‐based PGD of HEMA. Informative markers can also be easily co‐amplified with mutation‐containing F8 amplicons for combined mutation detection and linkage analysis.</abstract><cop>England</cop><pub>Elsevier Limited</pub><pmid>28345288</pmid><doi>10.1111/jth.13685</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Alleles Coagulation factors Diagnosis Dimorphism DNA Mutational Analysis Electrophoresis, Capillary Exons Factor VIII - genetics Factor VIII deficiency Female Fertilization in Vitro Gene loci Genetic Linkage Genetic Markers Genetic screening Genotype Haplotypes Hemophilia hemophilia A Hemophilia A - blood Hemophilia A - diagnosis Hemophilia A - genetics Heterozygote Humans In vitro fertilization INDEL Mutation Inversion Linkage analysis Male microsatellite Microsatellite Repeats Microsatellites Mutation Oocytes - metabolism Point Mutation Polymerase chain reaction Polymorphism, Genetic Pregnancy Preimplantation Diagnosis preimplantation genetic diagnosis Sequence Analysis, DNA short tandem repeat |
| title | Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A |
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