Ohtaki Hiroyuki, Yamamoto Toshimichi, Yoshimoto Takashi, Uchihi Rieko, Ooshima Chiaki, Katsumata Yoshinao, Tokunaga Katsushi
Department of Legal Medicine and Bioethics, Nagoya University, Nagoya, Japan.
Electrophoresis. 2002 Sep;23(19):3332-40. doi: 10.1002/1522-2683(200210)23:19<3332::AID-ELPS3332>3.0.CO;2-8.
A new multiplex system for six tetranucleotide short tandem repeat (STR) loci was devised based on multicolor dye technology. Six loci (D20S480, D6S2439, D6S1056, D9S1118, D4S2639, and D17S1290), each with high discriminating power (each unbiased estimates of expected heterozygosity, Exp. Hz, > 0.80 in a preliminary test), were selected from more than 100 tetranucleotide STRs included in a commercially available primer set. These loci were also selected so as not to link with general STRs in commercially released kits including the combined DNA index system (CODIS) 13 core STRs. The primers were newly designed in the present study, in which each amplicon size had a range of less than 200 base pairs (bp), in order to genotype from highly degraded template DNA. Using this system, we genotyped 270 Honshu (mainland)-Japanese and 187 Okinawa-Japanese. From these allele frequencies, we performed three tests, a homozygosity test, a likelihood ratio test and an exact test for Hardy-Weinberg equilibrium (HWE), and no significant deviations (p < 0.05) from HWE were observed. We also compared the allele distributions at six STRs between both populations, and they were significantly different (p < 0.05) at three loci (D6S2439, D9S1118 and D4S2639). Furthermore, the Exp. Hz and the power of discrimination (PD) at all loci in the Honshu-Japanese population were higher than 0.80 and 0.93, respectively. These statistical values for discriminating power in the Honshu-Japanese were almost the same as in the Okinawa-Japanese. This novel, multiplex polymerase chain reaction (PCR) amplification and typing system for six STR loci thus promises to be a convenient and informative new DNA profiling system in the forensic field.
基于多色染料技术，设计了一种用于六个四核苷酸短串联重复序列（STR）位点的新型多重系统。从市售引物组中包含的100多个四核苷酸STR中选择了六个位点（D20S480、D6S2439、D6S1056、D9S1118、D4S2639和D17S1290），每个位点都具有高鉴别力（在初步测试中，每个预期杂合度的无偏估计值，Exp. Hz，> 0.80）。选择这些位点也是为了使其不与市售试剂盒中的一般STR连锁，包括联合DNA索引系统（CODIS）的13个核心STR。在本研究中重新设计了引物，其中每个扩增子大小范围小于200个碱基对（bp），以便从高度降解的模板DNA中进行基因分型。使用该系统，我们对270名本州（日本大陆）人和187名冲绳人进行了基因分型。根据这些等位基因频率，我们进行了三项测试，即纯合性测试、似然比测试和哈迪-温伯格平衡（HWE）的精确测试，未观察到与HWE有显著偏差（p < 0.05）。我们还比较了两个群体在六个STR位点的等位基因分布，在三个位点（D6S2439、D9S1118和D4S2639）它们有显著差异（p < 0.05）。此外，本州人群中所有位点的Exp. Hz和鉴别力（PD）分别高于0.80和0.93。本州日本人的这些鉴别力统计值与冲绳日本人几乎相同。因此，这种用于六个STR位点的新型多重聚合酶链反应（PCR）扩增和分型系统有望成为法医领域一种方便且信息丰富的新DNA分型系统。
