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目的利用DNA条形码技术对中山口岸从柬埔寨进口木薯干中截获的外部形态特征破坏严重的2只未知鼠样本进行种类鉴定。方法提取截获鼠样本的基因组DNA,采用动物线粒体细胞色素氧化酶Ⅰ(COⅠ)的通用引物LCO1490和HCO2198扩增目的片段。然后测序、序列分析,与NCBI中的序列进行比对,并选取NCBI中序列相似度高的部分序列构建NJ树。结果 2只鼠的DNA条形码序列相似度高达99.8%,与NCBI中褐小家鼠(Mus cervicolor)的序列相似性最高为98.5%;大于DNA条形码判断物种的相似度98%的阈值。经科技查新,确认为全国口岸首次截获的国内未见分布鼠种。结论利用DNA条形码技术对截获鼠类进行鉴定,鉴定结果为褐小家鼠,为国内未见分布种。DNA条形码能克服传统物种鉴定依靠完整的外部形态的局限,并能克服鉴定资料、专业人员缺乏等因素带来的困难,有效提高外来物种的种类鉴别力和准确度。
OBJECTIVE: To identify the 2 unknown rat samples with the severely damaged external morphological features intercepted from the tapioca kernels imported from Cambodia by the DNA barcode technology. Methods The genomic DNA of the intercepted mouse samples was extracted and the target fragment was amplified using the universal primers LCO1490 and HCO2198 of animal mitochondrial cytochrome oxidase Ⅰ (COⅠ). Then sequenced and sequenced, and compared with the sequences in NCBI, NJN was constructed by selecting partial sequences with high similarity in NCBI. Results The similarity of the DNA barcode sequences of the two mice was 99.8%, the highest similarity was 98.5% with the sequence of Musca brasiliensis in NCBI, and the similarity was 98% of the DNA barcodes. The new search technology, confirmed as the first time the national ports intercepted no species of domestic distribution. Conclusion The identification of intercepted rodents using DNA barcoding technique showed that the results showed that there was no species in domestic. DNA barcodes can overcome the limitations of traditional species identification relying on complete external morphology and can overcome the difficulties caused by the identification of information, lack of professionals and other factors, and effectively improve the species discrimination and accuracy of alien species.