解旋恒温基因扩增技术在螨虫鉴定中的应用前景

Application prospect of helicase-dependent isothemal DNA amplification in identification of mites

中文关键词:  解旋恒温基因扩增  螨虫  鉴定

英文关键词:helicase dependent amplification  mites  identification

中文摘要:

      螨虫种类多, 分布广, 危害多样, 已成为人们关注的对象。随着进出口植物、植物产品和食品的增加, 其携带螨虫的风险越来越高, 许多国家把螨虫作为检疫对象设置技术壁垒。由于螨虫体型微小, 形态鉴定比较困难, 一直是口岸检疫的一大难点。随着现代分子技术的发展, 迫切需要建立快速、灵敏、准确的螨类检测方法。解旋酶恒温基因扩增技术(HDA)是一种简便、快速、高效、灵敏度高的体外恒温基因扩增技术。该技术依靠DNA解旋酶解开双链DNA、单链DNA结合蛋白(SSB)维持单链状态、DNA聚合酶催化靶片段的扩增。HDA不需要昂贵的仪器设备, 适用于基层实验室。HAD能扩增微生物基因组DNA、病原菌DNA、质粒DNA和cDNA等, 从其灵敏性、准确性和可操作性几方面来看, 该方法适用于螨虫鉴定。

英文摘要:

      The mite has become the focus of attention because of a large number of species, wide distribution and various hazards. The risk of carrying mites is getting higher and higher, with the increase of import and export of plants, plant products and food. Many countries have set up technical barriers by taking mites as quarantine pests. It is very difficult to identify the tiny mites by their morphological characteristics, so mite identification is one of the difficulties in quarantine. With the development of modern molecular technology, there is an urgent need to establish a rapid, sensitive, and accurate detection method of the mites in the quarantine work. The helicase-dependent amplification is a simple, rapid, efficient and sensitive isothermal gene amplification technology in vitro. It utilizes the action of helicases to separate double-stranded DNA, the single –stranded DNA-binding protein to maintain the single-stranded state and DNA polymerases to synthesize target fragments. The method does not require expensive equipments, and is very suitable for primary laboratory. It is reported that HDA can be used for the amplifications of microbial genomic DNA, microbial DNA, plasmid DNA and complementary DNA. From the sensitivity, accuracy and operability, HAD technology is suitable for the identification of the mites.