应用HRM技术进行法医昆虫种属鉴定的初步研究

目的对高分辨率熔解曲线技术(High Resolution Melting,HRM)在昆虫种属鉴定中的有效性进行初步研究。方法提取绿蝇头部、胸部、腿部组织基因组DNA,设计特异性引物对其线粒体COI和16SrDNA区进行扩增检测,结果分别用HRM和DNA测序两种方法进行检测,对其结果进行比较。结果 HRM方法对COI和16SrDNA基因检测均获得了检测峰,DNA测序COI基因区测序成功,16SrDNA基因区因扩增量少未能成功测序;两种方法对实验样本的胸部、腿部和头部组织的检测结果具有较好的一致性。结论 HRM和测序两种方法具有较好的一致性,可用于法医昆虫的种属鉴定研究。

Preliminary study of species identification by HRM in forensic entomology

Objective To explore the validity of High Resolution Melting（HRM）to be used for species identification in forensic entomology. Methods DNA was extracted from heads,throats and legs of flies,specified primers were designed to detect COI and 16SrDNA regions;after amplification,their products were detected both by HRM and DNA sequence. Results COI and 16SrDNA regions were detected successfully by HRM, COI was detected by DNA sequencing successfully, 16SrDNA was unsuccessfully due to insufficient amplified DNA sample, and the detective result of HRM is in accordance with the result of DNA sequencing in tissues from different parts of flies. Conclusions The HRM method has good consistency with DNA sequencing and could be used for species identification in forensic entomology.