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Accurate and Rapid Detection of Heterozygous Carriers of a Deletion by Combined Polymerase Chain Reaction and High-Performance Liquid Chromatography

We have developed a technique to detect accurately heterozygous carriers of a deletion. Specific target sequences were amplified by the polymerase chain reaction (PCR), and the products subsequently were analyzed by high-performance liquid chromatography. Examples from four loci demonstrated that 24...

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Published in:	Proceedings of the National Academy of Sciences - PNAS 1992-10, Vol.89 (19), p.9126-9130
Main Authors:	Asakawa, Jun-Ichi, Satoh, Chiyoko, Yamasaki, Yosuke, Chen, Shi-Han
Format:	Article
Language:	English
Subjects:	Alleles Base Sequence carriers Chromatography, High Pressure Liquid - methods Deoxyribonucleic acid detection DNA DNA - genetics Duchenne muscular dystrophy Duchenne's muscular dystrophy Exons Female Gene Deletion Genetic Carrier Screening Genetic loci Genetics Genomics Haptoglobins - genetics Hemoglobins - genetics Hemophilia A - genetics Hemophilia B heterozygosity high-performance liquid chromatography Homozygotes Humans Male man Molecular Sequence Data Mothers Oligodeoxyribonucleotides Polymerase chain reaction Polymerase Chain Reaction - methods Scientific imaging
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cited_by	cdi_FETCH-LOGICAL-c586t-af3f65419a4c7d0c16b8753d44ca6170c5f128e978e375b6f9cd84761b1dceba3
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container_issue	19
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container_title	Proceedings of the National Academy of Sciences - PNAS
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creator	Asakawa, Jun-Ichi Satoh, Chiyoko Yamasaki, Yosuke Chen, Shi-Han
description	We have developed a technique to detect accurately heterozygous carriers of a deletion. Specific target sequences were amplified by the polymerase chain reaction (PCR), and the products subsequently were analyzed by high-performance liquid chromatography. Examples from four loci demonstrated that 24-27 cycles of amplification for a single-copy DNA, based on 50 ng of genomic DNA, results in excellent quantitation that readily permits the detection of heterozygous carriers of a deletion. We have demonstrated that triplex PCR (three targets in a single PCR) entails no loss of precision. We also have demonstrated that this method can accurately differentiate the heterozygous carriers of a deletion from normal individuals in four family studies, three for Duchenne muscular dystrophy patients and one for a hemophilia B patient.
doi_str_mv	10.1073/pnas.89.19.9126
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Specific target sequences were amplified by the polymerase chain reaction (PCR), and the products subsequently were analyzed by high-performance liquid chromatography. Examples from four loci demonstrated that 24-27 cycles of amplification for a single-copy DNA, based on 50 ng of genomic DNA, results in excellent quantitation that readily permits the detection of heterozygous carriers of a deletion. We have demonstrated that triplex PCR (three targets in a single PCR) entails no loss of precision. 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Satoh, Chiyoko ; Yamasaki, Yosuke ; Chen, Shi-Han</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c586t-af3f65419a4c7d0c16b8753d44ca6170c5f128e978e375b6f9cd84761b1dceba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Alleles</topic><topic>Base Sequence</topic><topic>carriers</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Deoxyribonucleic acid</topic><topic>detection</topic><topic>DNA</topic><topic>DNA - genetics</topic><topic>Duchenne muscular dystrophy</topic><topic>Duchenne's muscular dystrophy</topic><topic>Exons</topic><topic>Female</topic><topic>Gene Deletion</topic><topic>Genetic Carrier Screening</topic><topic>Genetic loci</topic><topic>Genetics</topic><topic>Genomics</topic><topic>Haptoglobins - genetics</topic><topic>Hemoglobins - genetics</topic><topic>Hemophilia A - genetics</topic><topic>Hemophilia B</topic><topic>heterozygosity</topic><topic>high-performance liquid chromatography</topic><topic>Homozygotes</topic><topic>Humans</topic><topic>Male</topic><topic>man</topic><topic>Molecular Sequence Data</topic><topic>Mothers</topic><topic>Oligodeoxyribonucleotides</topic><topic>Polymerase chain reaction</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Scientific imaging</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Asakawa, Jun-Ichi</creatorcontrib><creatorcontrib>Satoh, Chiyoko</creatorcontrib><creatorcontrib>Yamasaki, Yosuke</creatorcontrib><creatorcontrib>Chen, Shi-Han</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Human Genome Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Asakawa, Jun-Ichi</au><au>Satoh, Chiyoko</au><au>Yamasaki, Yosuke</au><au>Chen, Shi-Han</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Accurate and Rapid Detection of Heterozygous Carriers of a Deletion by Combined Polymerase Chain Reaction and High-Performance Liquid Chromatography</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1992-10-01</date><risdate>1992</risdate><volume>89</volume><issue>19</issue><spage>9126</spage><epage>9130</epage><pages>9126-9130</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>We have developed a technique to detect accurately heterozygous carriers of a deletion. Specific target sequences were amplified by the polymerase chain reaction (PCR), and the products subsequently were analyzed by high-performance liquid chromatography. Examples from four loci demonstrated that 24-27 cycles of amplification for a single-copy DNA, based on 50 ng of genomic DNA, results in excellent quantitation that readily permits the detection of heterozygous carriers of a deletion. We have demonstrated that triplex PCR (three targets in a single PCR) entails no loss of precision. We also have demonstrated that this method can accurately differentiate the heterozygous carriers of a deletion from normal individuals in four family studies, three for Duchenne muscular dystrophy patients and one for a hemophilia B patient.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>1409613</pmid><doi>10.1073/pnas.89.19.9126</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record>
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source	PubMed (Medline); JSTOR Archival Journals and Primary Sources Collection【Remote access available】
subjects	Alleles Base Sequence carriers Chromatography, High Pressure Liquid - methods Deoxyribonucleic acid detection DNA DNA - genetics Duchenne muscular dystrophy Duchenne's muscular dystrophy Exons Female Gene Deletion Genetic Carrier Screening Genetic loci Genetics Genomics Haptoglobins - genetics Hemoglobins - genetics Hemophilia A - genetics Hemophilia B heterozygosity high-performance liquid chromatography Homozygotes Humans Male man Molecular Sequence Data Mothers Oligodeoxyribonucleotides Polymerase chain reaction Polymerase Chain Reaction - methods Scientific imaging
title	Accurate and Rapid Detection of Heterozygous Carriers of a Deletion by Combined Polymerase Chain Reaction and High-Performance Liquid Chromatography
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