新疆新地乡部分地区亚洲璃眼蜱的形态及分子生物学鉴定

Morphological classification and molecular identification of Hyalomma asiaticum in parts of Xindi Township,Xinjiang

目的了解新疆裕民县新地乡所采集蜱种的形态学及分子生物学情况。方法采集该地区家畜体表寄生蜱,借助超景深三维显微系统VHX 600对收集到的蜱进行初步的形态学鉴定。提取全蜱总DNA,采用PCR技术,基于COI和ITS2基因位点对蜱DNA进行扩增,将阳性的PCR产物进行测序。通过NCBI数据库中Blast在线分析软件,将测序所获序列与数据库中其他相关序列进行同源性比对分析。利用MEGA 7.0软件中的邻接法构建遗传进化树,确定蜱的进化生物学特性。结果经形态学鉴定,从裕民县新地乡采集的蜱均符合亚洲璃眼蜱的特征。基于COI和ITS2基因序列建立的遗传进化树表明,本次研究所采集的蜱与已知的亚洲璃眼蜱聚为一支。结论通过形态学及分子生物学鉴定,确定本次研究所采集的蜱为亚洲璃眼蜱。该结果为新疆新地乡蜱种分布提供一定数据资料,为蜱种鉴定和遗传进化特征分析提供参考。

The purpose of this study was to identify the tick species native to Xindi Township,Yumin County,Xinjiang,China.Preliminary morphological identification of parasitic ticks collected from animals in the area was conducted with an ultra-depth of field three-dimensional VHX 600 digital stereo microscope.Total DNA of the ticks was extracted,amplified by PCR based on the COI and ITS2 gene loci,and the positive PCR products were sequenced.The sequence were aligned with reference sequences from the NCBI database were aligned with the Basic Local Alignment Search Tool.A genetic phylogenetic tree was generated with the neighbor-joining method of MEGA 7.0 software to determine the evolutionary biological characteristics of ticks.Morphological identification showed that the ticks collected from Xindi Township of Yumin County were consistent with the characteristics of Hyalomma asiaticum.An evolutionary tree based on the COI and ITS2 gene sequences showed that the ticks collected in this study were clustered with known H.asiaticum sequences.The PCR products of COI and ITS2 were sequenced and compared,which confirmed that the collected tick species were H.asiaticum,in agreement with the morphological and molecular biological results.These findings help to clarify the distribution of ticks in Xindi Township of Xinjiang,and provide basic data for the analysis of tick genetic and evolutionary characteristics,as reference for surveillance and control of ticks in the Xinjiang Uygur Autonomous Region.