Abstract：
Domestic dogs are helpers in outdoor human work and companions for families; thus, individual canine identification and parentage testing are crucial in certain fields, including forensics and breeding programs. In this study, a six-dye fluorescent labeling multiplex amplification system containing 29 canine short tandem repeats (STRs) and the sex-determining marker DAmel was developed. The system was called the Tronfo Canine 30-plex STR Kit and was further validated according to the Scientific Working Group on DNA Analysis Methods and the Organization of Scientific Area Committees for Wildlife Forensics guidelines, including tests for PCR conditions, precision, species specificity, sensitivity, stability, repeatability and reproducibility, a population study, and a study of 16 paternity test cases. The results indicated that the novel canine STR assay was accurate, specific, reproducible, stable, and robust. Complete profiles were obtained with 31.25 pg of canine DNA. Additionally, 500 unrelated canine individuals were investigated using this novel system, and the combined power of discrimination and exclusion values were 0.999999999999999999 and 0.999996451039850, respectively. These results suggest that the Tronfo Canine 30-plex STR Kit is highly polymorphic, informative, and suitable for individual canine identification and parentage testing.
摘要：
家犬是户外人类工作的助手和家庭的伴侣；因此，在某些领域，个体犬的识别和亲子关系测试至关重要，包括法医和育种计划。在这项研究中,开发了包含29个犬短串联重复序列（STR）和性别决定标记DAmel的六染料荧光标记多重扩增系统。该系统被称为Tronfo犬30-plexSTR试剂盒，并根据DNA分析方法科学工作组和野生动物取证科学领域委员会组织指南进行了进一步验证，包括PCR条件的测试，精度,物种特异性，灵敏度,稳定性,重复性和再现性，一项人口研究,以及对16例亲子鉴定案例的研究。结果表明，新的犬STR检测方法是准确的，具体,可重复,稳定,和强大的。用31.25yg的犬DNA获得完整的图谱。此外,使用这个新颖的系统调查了500个无关的犬类个体，判别力和排除力的综合值分别为0.99999999999999999999999999999451039850。这些结果表明，Tronfo犬30-plexSTR试剂盒是高度多态的，翔实，适用于个体犬类鉴定和亲子鉴定。
