van Asch Barbara, Alves Cíntia, Gusmão Leonor, Pereira Vânia, Pereira Filipe, Amorim António
IPATIMUP - Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Portugal.
Electrophoresis. 2009 Jan;30(2):417-23. doi: 10.1002/elps.200800307.
A single multiplex PCR assay capable of simultaneously amplifying nine canine-specific autosomal STR markers (FH3210, FH3241, FH2004, FH2658, FH4012, REN214L11, FH2010, FH2361 and the newly described C38) was developed for individual identification and parentage testing in domestic dogs. In order to increase genotyping efficiency, amplicon sizes were optimized for a 90-350 bp range, with fluorescently labelled primers for use in Applied Biosystems, Inc., platforms. The performance of this new multiplex system was tested in 113 individuals from a case-study population and 12 random dogs from mixed-breed origin. Co-dominant inheritance of STR alleles was investigated in 101 father, mother and son trios. Expected heterozygosity values vary between 0.5648 for REN214L11 and 0.9050 for C38. The high level of genetic diversity observed for most markers provides this multiplex with a very high discriminating power (matching probability=1.63/10(10) and matching probability among siblings=4.9/10(3)). Allele sequences and a proposal for standardized nomenclature are also herein presented, aiming at implementing the use of this system in forensic DNA typing and population genetic studies. This approach resulted in an optimized and well-characterized canine DNA genotyping system that is highly performing and straightforward to integrate and employ routinely. Although this STR multiplex was developed for use and tested in a case-study population, the Portuguese breed Cão de Gado Transmontano, it proved to be useful for general identification purposes or parentage testing.
开发了一种单一的多重聚合酶链反应（PCR）检测方法，能够同时扩增9个犬特异性常染色体短串联重复序列（STR）标记（FH3210、FH3241、FH2004、FH2658、FH4012、REN214L11、FH2010、FH2361和新描述的C38），用于家犬的个体识别和亲子鉴定。为了提高基因分型效率，将扩增子大小优化到90 - 350 bp范围，并使用荧光标记引物，以用于应用生物系统公司的平台。在一个案例研究群体的113个个体和12只混种来源的随机犬中测试了这种新的多重系统的性能。在101个父、母和子三人组中研究了STR等位基因的共显性遗传。预期杂合度值在REN214L11的0.5648和C38的0.9050之间变化。大多数标记观察到的高水平遗传多样性为这种多重检测提供了非常高的鉴别能力（匹配概率 = 1.63 / 10(10)，同胞间匹配概率 = 4.9 / 10(3)）。本文还给出了等位基因序列和标准化命名法的建议，旨在将该系统应用于法医DNA分型和群体遗传学研究。这种方法产生了一个经过优化且特征明确的犬DNA基因分型系统，该系统性能高，易于整合且常规使用。尽管这种STR多重检测是为在一个案例研究群体——葡萄牙品种Transmontano役用犬中使用和测试而开发的，但它被证明可用于一般识别目的或亲子鉴定。
