A new 39-plex analysis method for SNPs including 15 blood group loci

A novel 39-plex typing system for single nucleotide polymorphisms (SNPs) has been developed. This multiplex approach has the advantage of being able to type 38 autosomal SNPs and one sex-discriminating base exchange site on the X and Y chromosomes rapidly and simultaneously. The SNP loci on the auto...

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Veröffentlicht in:Forensic science international 2004-08, Vol.144 (1), p.45-57
Hauptverfasser: Inagaki, Sachiyo, Yamamoto, Yuji, Doi, Yusuke, Takata, Tomoyo, Ishikawa, Takaki, Imabayashi, Kiyomi, Yoshitome, Kei, Miyaishi, Satoru, Ishizu, Hideo
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container_issue 1
container_start_page 45
container_title Forensic science international
container_volume 144
creator Inagaki, Sachiyo
Yamamoto, Yuji
Doi, Yusuke
Takata, Tomoyo
Ishikawa, Takaki
Imabayashi, Kiyomi
Yoshitome, Kei
Miyaishi, Satoru
Ishizu, Hideo
description A novel 39-plex typing system for single nucleotide polymorphisms (SNPs) has been developed. This multiplex approach has the advantage of being able to type 38 autosomal SNPs and one sex-discriminating base exchange site on the X and Y chromosomes rapidly and simultaneously. The SNP loci on the autosomes, which we examined, contain 15 loci distributed on blood type genes: three on RhCE, two each on Km and Gc, and one each on Duffy, AcP1, Tf, MN, GPT, EsD, PI, and Kidd genes. Thirty-seven genomic DNA fragments containing a total of 38 SNPs and one sex-discriminating site were amplified in one multiplex PCR reaction. Following the reaction, single nucleotide primer extension reaction was performed by dividing these SNP loci into five groups. The SNP type of each of the 39 loci was determined at one time by capillary electrophoresis using the newly designed multi-injection method. The combined PD (power of discrimination) of this typing system was (1–1.1)×10 −14, and the MEC (mean exclusion chance) was 0.9990. We applied this system to forensic cases, including 16 paternity testing cases (13 non-exclusion and three exclusion cases) and one personal identification case. For the paternity testing cases, the highest Essen–Möller’s W-value was 0.9999995. The pM (matching probability) of the personal identification case was 2.22×10 −17. These data showed that this system was an excellent tool for use in forensic cases of paternity testing and personal identification.
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This multiplex approach has the advantage of being able to type 38 autosomal SNPs and one sex-discriminating base exchange site on the X and Y chromosomes rapidly and simultaneously. The SNP loci on the autosomes, which we examined, contain 15 loci distributed on blood type genes: three on RhCE, two each on Km and Gc, and one each on Duffy, AcP1, Tf, MN, GPT, EsD, PI, and Kidd genes. Thirty-seven genomic DNA fragments containing a total of 38 SNPs and one sex-discriminating site were amplified in one multiplex PCR reaction. Following the reaction, single nucleotide primer extension reaction was performed by dividing these SNP loci into five groups. The SNP type of each of the 39 loci was determined at one time by capillary electrophoresis using the newly designed multi-injection method. The combined PD (power of discrimination) of this typing system was (1–1.1)×10 −14, and the MEC (mean exclusion chance) was 0.9990. We applied this system to forensic cases, including 16 paternity testing cases (13 non-exclusion and three exclusion cases) and one personal identification case. For the paternity testing cases, the highest Essen–Möller’s W-value was 0.9999995. The pM (matching probability) of the personal identification case was 2.22×10 −17. These data showed that this system was an excellent tool for use in forensic cases of paternity testing and personal identification.</abstract><cop>Kidlington</cop><pub>Elsevier Ireland Ltd</pub><pmid>15240020</pmid><doi>10.1016/j.forsciint.2004.03.005</doi><tpages>13</tpages></addata></record>
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subjects Analysis
Biological and medical sciences
Blood Group Antigens - genetics
Blood group loci
Blood groups
DNA Fingerprinting - methods
DNA Primers
Electrophoresis, Capillary
Female
Forensic genetics
Forensic sciences
Fundamental and applied biological sciences. Psychology
Gene Frequency
Genes
Genes. Genome
Genetics
Genomics
Humans
Identification
Investigative techniques, diagnostic techniques (general aspects)
Male
Medical sciences
Methods
Microbiology
Molecular and cellular biology
Molecular genetics
Mutation
Paternity
Paternity testing
Personal identification
Polymerase Chain Reaction
Polymorphism, Single Nucleotide
Power of discrimination
Sequence Analysis, DNA
Single nucleotide polymorphisms
Single nucleotide primer extension
Tandem Repeat Sequences
title A new 39-plex analysis method for SNPs including 15 blood group loci
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