用随机扩增多态DNA技术区分7种硬蜱  

DIFFERENTIATION OF SEVEN SPECIES OF TICKS BY RANDOM AMPLIFIED POLYMORPHIC DNA TECHNIQUE

作　　者：杨银书[1] 常德辉[2] 赵红斌[2] 史智勇[2] 马黎萍[1] 张继军[1] 第五进学[1] 

机构地区：[1]兰州军区军事医学研究所,甘肃兰州730020 [2]兰州军区总医院

出　　处：《中国寄生虫病防治杂志》2004年第5期301-303,共3页Chinese Journal of Parasitic Disease Control

基　　金：甘肃省中青年科研基金项目 (No .YS 0 0 1 A2 3 0 2 5)

摘　　要：目的　分析 7种硬蜱基因组DNA的随机扩增多态性 ,探讨随机扩增多态性DNA(RAPD)技术在硬蜱分类中的应用。　方法　提取草原革蜱、森林革蜱、青海血蜱、台湾血蜱、刻点血蜱、龟形花蜱、卵形硬蜱DNA。选取 3条不同的随机排列碱基顺序的多聚核苷酸单链为引物 (P5、P6、P7)进行RAPD PCR扩增反应。将扩增产物制备成DNA图谱 ,对这 7种硬蜱的DNA多态性进行分析。　结果　 7种硬蜱分别扩增出不同数量与不同分子质量的DNA片段。有些硬蜱基因组DNA扩增产物中具有相同的DNA条带 ,这反映出它们的基因组DNA具有同源性 ;同时又具有各自独特的DNA条带 ,且DNA条带的亮度也有差异。　结论　RAPD技术可以准确地区分这 7种蜱。Objective To analyze DNA polymorphisms of seven species of ticks and probe into the application of random amplified polymorphic DNA(RAPD) technique in identification of ticks. Methods Genome DNA was extracted respectively from Dermacentor nuttalli, Dermacentor silvarum, Haemaphysalis qinghaiensis, Haemaphysalis formosensis, Haemaphysalis punctata, Amblyomma testudinarium, Ixodes ovatus. Three arbitrary primers(P 5 , P 6 , P 7 ) were selected, the genome DNA of the seven species of ticks was respectively amplified by RAPD-PCR. The PCR products were analyzed on a 1.5% agarose gel. The DNA polymorphisms of the seven species of ticks were analyzed according to their fingerprint maps. Results The DNA maps that the numbers and molecular weight of DNA fragments were different among the seven species of ticks were gained by RAPD technique. The amplified products of genome DNA of some species of ticks had common fragments, which indicated that the genomes of these species of ticks had homogeneity to some degree. Meanwhile, the genomes of the seven species of ticks had their peculiar DNA fragments and the differences in the brightness of the DNA fragments existed. Conclusion The RAPD is an exact method for ticks taxonomy.

关 键 词：蜱 基因组DNA 分类 随机扩增多态性DNA(RAPD) 

分 类 号：R384.4[医药卫生—医学寄生虫学]