白纹伊蚊Cytb基因特异性引物的设计及评价  

Designation and evaluation of specific primers for Aedes albopictus

作　　者：王颖 王勇 孙超 段效辉 方绍庆 WANG Ying;WANG Yong;SUN Chao;DUAN Xiao-hui;FANG Shao-qing(Yantai International Travel Healthcare Center,Yantai,Shandong 264000,China;不详)

机构地区：[1]烟台国际旅行卫生保健中心,山东烟台264000 [2]临沂海关 [3]烟台海关

出　　处：《中国国境卫生检疫杂志》2020年第4期259-261,共3页Chinese Journal of Frontier Health and Quarantine

基　　金：原国家质检总局科技计划项目(2016IK313);原山东出入境检验检疫局科技计划项目(SK201719)。

摘　　要：目的设计并评价白纹伊蚊细胞色素b(Cytb)基因特异性引物。方法根据白纹伊蚊Cytb基因保守区域序列,设计1对引物,优化PCR扩增的退火温度。以白纹伊蚊及同属、同科、同目昆虫基因组DNA为样本,评价引物的特异性;以白纹伊蚊基因组DNA为样本,评价引物的灵敏性;扩增4个口岸捕获的8只白纹伊蚊,评价引物的重复性。结果设计的引物的最佳退火温度为55℃。该引物可成功扩增白纹伊蚊Cytb基因,不能扩增其同属、同种、同目昆虫基因;可扩增来自4个口岸的浓度1.9 ng/μl以上的白纹伊蚊基因组DNA。结论本研究设计的引物的特异性、灵敏性、重复性理想,适用于白纹伊蚊的鉴定。Objective To design and evaluate specific primers for Cytb gene of Aedes albopictus.Methods A pair of primer was designed based on the conserved region of Cytb gene of Aedes albopictus,then the annealing temperature was optimized.The genomic DNA of Aedes albopictus and medical vectors of the same genus,family and order were used as samples to evaluate the specificity of the primers.The repeatability was evaluated by amplifying the genomic DNA of eight Aedes albopictus from four ports in China.Results The optimum annealing temperature for the primers was 55℃.The primers successfully amplified the Cytb gene of Aedes albopictus,but not the genes of the same genus,species and order medical vectors.The genomic DNA of Aedes albopictus collected at 4 ports with a concentration of more than 1.9 ng/μl was amplified.Conclusion The primers designed in this study showed ideal specificity,sensitivity and repeatability for the identification of Aedes albopictus.

关 键 词：白纹伊蚊 细胞色素B基因 引物 

分 类 号：R185[医药卫生—流行病学]